(manual antropometría enasem final...

ESTUDIO NACIONAL DE SALUD Y ENVEJECIMIEN TO EN MÉXICO (ENASEM-II)

Manual de Procedimientos Antropométricos y Muestra Biológica

Septiembre del 2012

2012

InstitutoNacional de SaludPública Centro de Investigación en Evaluación y Encuestas


1

Estudio Nacional de Salud y Envejecimiento en México (ENASEM-II)

Responsable: Juan Pablo Gutiérrez ([email protected])

Elaboración

Responsable: Aurora Franco Núñez ([email protected])


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INDICE

Introducción ...................................................................................................................... 3

Aspectos Generales ........................................................................................................ 5

Funciones Y Responsabilidades del Antropometrista ..................................................... 5

Lineamientos Generales para el Levantamiento ............................................................. 8

Procedimientos para un Levantamiento de Calidad ........................................................ 8

Presión Arterial .............................................................................................................. 10

Puntos Antropométricos ................................................................................................ 12

La Antropometría ........................................................................................................... 12

Peso .............................................................................................................................. 13

Talla ............................................................................................................................... 16

Circunferencia de Cintura .............................................................................................. 19

Circunferencia de Cadera .............................................................................................. 21

Medición de la Altura de la Rodilla ................................................................................ 22

Medición de Altura Sentado ......................................................................................... 24

Medidas de Desempeño ................................................................................................ 25

Velocidad de la Marcha ................................................................................................. 27

Prueba de la Fuerza de Presión. ................................................................................... 30

Obtención de Muestras de Sangre Capilar .................................................................... 33

Determinación de Hemoglobina Glucosilada ................................................................. 34

Determinación de Hemoglobina .................................................................................... 38

Toma de Muestra Sanguínea ........................................................................................ 43

Técnica para Extracción Venosa ................................................................................... 45

Procedimiento Después de la Toma de Muestra Sanguínea ........................................ 48

Diagrama de Flujo de la Antropometría ......................................................................... 49

Bibliografía. .................................................................................................................... 50

Apéndice I. Biomarcadores ............................................................................................ 51

Apéndice II. Nota Técnica sobre el Método de Análisis de Laboratorio para la Determinación de Colesterol .......................................................................................... 67


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INTRODUCCION

El Estudio Nacional de Salud y Envejecimiento en México (ENASEM, II) es un estudio

prospectivo poblacional acerca de la dinámica de la salud y el envejecimiento en

México. El objetivo general del estudio es contribuir en la generación de nuevo

conocimiento sobre envejecimiento y salud de los adultos mayores en México.

El estudio incluye una encuesta longitudinal1 en hogares usando una muestra

representativa nacional de personas de 50 años o más, la cual se levantó en 2001 con

seguimiento a los mismos sujetos en 2003. El levantamiento actual de la ENASEM

representa la tercera medición del estudio.

Participan en el estudio hombres y mujeres de 50 años y más seleccionados en el

estudio desde el 2001, así como sus respectivos cónyuges. Para el 2012 se contará

con una muestra adicional de nuevos participantes. En el levantamiento de la

información participa el Instituto Nacional de Estadística Geografía e Informática

(INEGI) en la aplicación de los cuestionarios y el Instituto Nacional de Salud Pública

(INSP) en la realización de mediciones antropométricas, medidas de desempeño

funcional y en la toma de la tensión arterial. Así mismo, el INSP se encargara de la

aplicación de pruebas in sitúo para la determinación de hemoglobina y hemoglobina

glucosilada y de la obtención de una muestra de sangre para la determinación en

laboratorio de biomarcadores biológicos y otra muestra será almacenada para realizar

en un futuro estudios genéticos (Los factores genéticos se asocian con enfermedades

comunes como la diabetes y las enfermedades cardiovasculares, y se relacionan con

aspectos biológicos del proceso de envejecimiento).

Cabe mencionar que mientras el INEGI aplicará los cuestionarios al total de

participantes en el estudio, el INSP se enfocará en una submuestra de los mismos. La

sub-muestra estará comprendida por la muestra total de cuatro estados.

1 Un estudio de seguimiento es aquel en el que un grupo de sujetos usualmente seleccionados al azar

es medido en varias ocasiones el tiempo, es decir, se repite el estudio en varios momentos en el

tiempo.


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En estas áreas, el personal del INEGI llevará a cabo la entrevista en el hogar y

notificará a los encuestados que acepten participar en el estudio que en

aproximadamente dos semanas, el personal de salud del INSP acudirá al hogar a

solicitar su participación en una segunda fase del estudio.

El INEGI entregará periódicamente al INSP el listado de personas que aceptaron

responder los cuestionarios [participantes y cónyuges (si aplica)]. El personal del

INSP identificará a dichas personas y previo consentimiento informado se efectuarán

los siguientes procedimientos:

Mediciones antropométricas (talla, peso, circunferencia de cintura y cadera, talla

sentado y altura de la rodilla, tensión arterial)

Pruebas de desempeño [balance con el pie derecho e izquierdo, velocidad de la

marcha y fuerza de agarre].

Toma de muestras de sangre venosa

Toma de muestra de sangre capilar

En el presente manual se establecen los lineamientos para efectuar correctamente las

mediciones antropométricas, medidas de desempeño, toma de tensión arterial y toma

de muestras de sangre.


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Aspectos generales

Un adecuado levantamiento de información se basa en el cumplimiento de las

funciones y responsabilidades del antropometrista, en el seguimiento de los

procedimientos establecidos para el trabajo de campo y en el conocimiento y dominio

del programa de cómputo para la captura de la información, entre otros aspectos.

La calidad de la información obtenida y el éxito del proyecto se basan en un excelente desempeño en campo del

antropometrista.

Funciones y responsabilidades del antropometrista

La función principal del antropometrista es aplicar los procedimientos establecidos

en cada medición, en el 100% de los adultos seleccionados. Esta función se logra con

las siguientes acciones a seguir:

Garantizar la calidad de los procedimientos realizados y el completo llenado de la

información, exceptuando los casos en los que se indique la omisión de algunas

mediciones.

Emplear habilidades y estrategias de convencimiento para evitar el rechazo a realizar

las mediciones.

Recopilar toda la información requerida en el tiempo destinado para cada localidad.

En caso de ser necesario, hacer las visitas adicionales necesarias para evitar

ausencia de información.

Estar atento a todas las indicaciones que se le instruyan y ser puntual en todos sus

compromisos de trabajo.

Apoyar otros trabajos comisionados por el supervisor y participar en las reuniones de

revisión que éste organice.

Comunicar al supervisor los avances logrados, así como los percances o dificultades

que pudieran afectar el levantamiento.


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Reportar inmediatamente al supervisor toda situación anómala o irregular que se

presente durante el trabajo de campo.

Apoyar a sus compañeros de equipo para la conclusión de las actividades previstas

en cada vivienda.

Cuando por algún motivo las mediciones no puedan llevarse a cabo en algún

seleccionado, el antropometrista deberá solicitar la firma de algún integrante del hogar

en un documento donde se especifique la razón, mismo que entregará al supervisor

quien realizará la revisión y verificación correspondientes.

Si no encontrara a alguien en la vivienda y los vecinos o alguna otra persona le

comentaran que la casa está desocupada o que los propietarios están temporalmente

fuera de la localidad, deberá indicarlo en un documento escrito (puede ser en una

hoja en blanco o bien en su bitácora), especificando el nombre y el domicilio de la

persona que le proporcione esta información.

En caso de que requiera hacer aclaraciones durante el vaciado de información, el

sistema dispone de una sección de “Observaciones” que podrá abrir en cualquier

momento, para especificar claramente la causa o circunstancia por la que se realiza la

observación.

Por otra parte, la responsabilidad primordial del antropometrista es realizar cada

procedimiento con excelente calidad. Para lograr este objetivo se deben tener en

cuenta los siguientes principios:

Calidad. La excelente calidad está determinada fundamentalmente por la aplicación

correcta de las técnicas, procedimientos y la cobertura alcanzada de acuerdo con la

selección de la muestra.


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Productividad. Es de suma importancia el cumplimiento de los estándares de

productividad del levantamiento. Esto requiere la toma total de las mediciones en el

tiempo establecido; de ahí la necesidad de cumplir con las cargas de trabajo en los

periodos acordados.

Confidencialidad. Se debe guardar estricta confidencialidad sobre la información

obtenida en cada hogar. A través de la carta de consentimiento los antropometristas

notificaran a los informantes que la información obtenida no será revelada a otras

personas.2

Respeto. El entrevistador debe mostrar respeto en todo momento hacia las

tradiciones de la zona y hacia los diversos grupos de personas que habitan en ella.

Otra de las responsabilidades de suma importancia del antropometrista es devolver

todos los materiales de trabajo al finalizar el operativo de campo, principalmente de

la computadora portátil (PC Laptop) que le haya sido asignada, así como el equipo

para cada medición (Estadímetro, Báscula, y Hemocue, etc.). En caso contrario, se

deberá presentar por escrito la justificación correspondiente, el acta administrativa, el

acta judicial o realizar el pago respectivo.

2 Conforme a las disposiciones del Artículo 38 de la Ley de Información Estadística y Geográfica en

vigor, “Los datos e informes que los particulares proporcionen para fines estadísticos o provengan de

registros administrativos o civiles, serán manejados, para efectos de esta LEY, bajo la observancia de

los principios de confidencialidad y reserva y no podrán comunicarse, en ningún caso, en forma

normativa o individualizada, ni harán prueba ante autoridad administrativa o fiscal, ni en juicio o fuera

de él.”


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Lineamientos generales para el levantamiento

Procedimientos para un levantamiento de calidad

La recopilación de la información de las mediciones antropométricas y biológicas

se realizará mediante la aplicación de los procedimientos a los adultos de 50 años y

más seleccionados en el estudio y de ser el caso de sus respectivos cónyuges. Para

lograr un resultado de calidad, es importante que los antropometristas cumplan con lo

siguiente:

Tener claros los objetivos del levantamiento. Es común que las personas

entrevistadas pidan información acerca de lo que se busca con las mediciones, por lo

que antes de salir a campo es necesario conocer los antecedentes conceptuales del

proyecto y resolver cualquier duda al respecto.

Realizar las mediciones con los informantes e ingresar los datos cuidadosamente

en el programa de cómputo.

Nota: Es sumamente importante realizar las mediciones con toda la privacidad

posible, pues la presencia de otras personas puede influir en el informante y, en

consecuencia, se corre el riesgo de no obtener las mediciones.

Mostrar la credencial de acreditación como personal del Instituto Nacional de

Salud Pública (INSP) y portarla en un lugar visible. Esta acción representa un

respaldo para la confianza del adulto.

Propiciar un ambiente favorable. Existe una gran cantidad de grupos humanos que

tienen diversas maneras de concebir y organizar su vida. Este hecho, asociado con

las diferentes personalidades de cada individuo, obligan a poner en práctica toda la

habilidad y sensibilidad del antropometrista para establecer un ambiente de confianza

y privacidad durante la toma de las mediciones.


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Todos los antropometristas deben seguir uniformemente los anteriores

procedimientos , de esta manera, se logrará la homogeneidad en el trabajo de

campo, característica básica para que éste adquiera validez y que la información

obtenida pueda ser analizada en su conjunto y con un alto nivel de veracidad y

precisión.


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Presión arterial

� Concepto

La Presión Arterial (PA) se refiere a la fuerza que produce la sangre sobre las

arterias, al pasar por ellas. Las arterias son vasos sanguíneos que llevan sangre

desde el corazón hacia todo el cuerpo. La sangre a su vez transporta el oxigeno y los

nutrientes a todos los órganos del cuerpo para que puedan funcionar

La presión arterial se compone de dos cifras:

La máxima o sistólica que es cuando el corazón bombea la sangre.

La mínima o diastólica en el momento que el corazón se relaja.

La presión alta o hipertensión arterial se define como una elevación continua o

intermitente de la presión de la sangre, ya sea diastólica o sistólica. Cuando la presión

arterial es alta, empieza a dañar los vasos sanguíneos, el corazón y los riñones. Esto

puede provocar un ataque al corazón, un ataque cerebral, enfermedades de los

riñones y otros problemas.

� Cifras normales

Se considera como valor normal 120/80 mm/hg en adultos. Sin embargo, estas cifras

pueden variar dependiendo de la constitución física, edad y sexo del individuo, por lo

que para considerar una presión normal debemos preguntar a la persona si conoce el

valor de su presión arterial, puesto que algunos mantienen presiones bajas sin tener

ningún problema. Se considera hipertensión arterial cuando la presión sistólica es

mayor a 140 mm de Hg y la presión diastólica es mayor de 90 mm de Hg.


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� Equipo y material

Baumanómetro electrónico (OMRON)

Pilas AA

Lápiz o pluma

Bitácora de registro

Tarjeta de resultados

� Técnica de Medición

• Se realizarán dos mediciones.

• Preparar el baumanómetro, es decir instalar la manguera del brazalete al

baumanómetro del lado izquierdo y encender el botón azul de ON/OFF.

• En la parte inferior izquierda de la pantalla aparece el nombre Systolic

(Sistólica) y Diastolic (Diastólica), al encender la pantalla aparece con la cifra

688 en cada una de ellas y en el apartado de pulso aparece la cifra 188 y el

reloj marcador del tiempo

• Explicar al adulto el procedimiento que se le va a realizar.

• Pedir al adulto que se siente y que se descubra el brazo izquierdo, en caso que

no pueda hacerlo solo, ayúdelo.

• Pedirle que se retire, anillos, reloj, pulseras etc.

• Debe ponerse cómodo, en posición sentado en una silla con brazos o junto a

una mesa, de tal manera que permita colocar el brazo bien extendido y a la vez

mantenerlo apoyado para facilitar la medición.

• Localizar el pulso humeral con los dedos índice y medio, ajustar el brazalete de

tal manera que la manguera no se obstruya y quede sobre el trayecto de la

arteria.

• Colocar el brazalete alrededor del brazo, a dos centímetros por arriba del codo.

• Verifique que el brazalete tenga contacto con la piel sin apretar.


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• Una vez colocado el brazalete correctamente, se oprime el botón gris de

START el insuflará el brazalete y una vez que deje de descender la

numeración, inmediatamente después aparecerá el valor, mismo que deberá

registrarlo.

• Retirar el brazalete del brazo del adulto.

Nota: Antropometrista recuerde que para realizar la segunda toma de presión arterial,

el adulto tiene que permanecer sentado por 5 minutos.

Baumanómetro digital

Puntos antropométricos

La antropometría

La antropometría es una técnica amigable y poco costosa, portátil y aplicable en todo

el mundo para evaluar el tamaño, las proporciones y la composición del cuerpo

humano. Refleja el estado nutricional y de salud y permite predecir el rendimiento, la

salud y la supervivencia. Como tal, es un instrumento valioso en la orientación de las

políticas de salud pública y las decisiones clínicas.


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Es la técnica sistematizada de medir y realizar observaciones en el cuerpo humano,

en el esqueleto, que describe las diferencias cuantitativas de las medidas del cuerpo

utilizando métodos adecuados y científicos. La amplitud de las observaciones y

medidas está limitada únicamente por la naturaleza de los problemas a los cuales se

aplica; en consecuencia, las reglas, divisiones, medidas e índices tienen en todo

momento carácter “convencional”.

Son las maniobras en las cuales se obtiene el peso en kilogramos, la talla de pie, la

talla sentado y las circunferencias de cintura, cadera y talón-rodilla en centímetros,

por mencionar algunas.

Nota general

Los puntos antropométricos que a continuación se describen serán realizados 2

veces, en caso necesario de duda o margen de error se realizara una 3ra medición.

Peso

� Concepto

Es la resultante de la acción que ejerce la gravedad sobre un cuerpo. (Las variaciones

que pueda haber en observación son debidas al sexo, edad del individuo y muchos

otros factores).

Es la medida de valoración nutricional más empleada, en concepto de peso, es un

indicador de masa corporal total necesaria para detectar en combinación con la talla

alteraciones en el estado nutricional tales como obesidad o desnutrición


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- Una báscula estándar

- Una tara de 5kg.

- Toallas de papel

- Lápiz o pluma

- Bitácora de registro

- Tarjeta de resultados

Para adultos se utilizarán básculas portátiles electrónicas digitales con precisión de

100g. El peso máximo que registra la báscula es de 150.0 Kg.

� El funcionamiento y calibración de la báscula deberá revisarse con la ayuda de

taras (5Kg.) para estar seguros de que va a dar pesos exactos.


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� Indicaciones generales para peso

Explicar al adulto el procedimiento que se le va realizar.

Pedirle que se quede con el mínimo de ropa aceptable y que se quite los zapatos y

objetos pesados que sobreestimen el peso como pueden ser: llaves, monedas,

cinturones con hebillas gruesas, chamarras o suéteres, chalecos pesados etc.

Evitar pesarlos con ropa pesada o húmeda, mujeres con cabello largo mojado.

Técnica de Medición

• Indicar antes de proceder a pesar, si tienen el pantalón muy largo pedirle o

realizarle el doblez hacia arriba, de tal manera que pueda observar los talones

y punta de los pies de la persona los cuales deben de permanecer dentro de

los bordes de la bascula y tener buena visión de la pantalla.

• Cuando sea posible, evacuar la vejiga. Nunca debe pesarse después de haber

realizado una comida abundante.

• Pedir al adulto que de un solo paso se suba a la báscula como lo indica la

imagen siguiente.

Nota

Sólo se ayudará a subir a la báscula a los adultos con alguna discapacidad.

En caso de que la iluminación sea insuficiente, utilice una lámpara de mano para

poder observar el resultado.


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Talla

� Concepto

Es la distancia tomada en posición vertical, de pie desde el suelo al vértex o punto

más alto del cráneo.

El estado enfermizo o saludable de los individuos está en íntima relación con la talla,

siendo los principales factores la genética y la nutrición.


- Estadímetro

- Lápiz o pluma

- Sanitas


- Tarjeta de resultados.


• Explicar al individuo el procedimiento que se le va a realizar.

• Se busca dentro o fuera de la casa un lugar donde el piso se encuentre lo más

plano posible, debe tener un ángulo de 90°, se colo ca la base del estadímetro

para ensamblar las partes de la regleta graduada.

• Verificar que al unir las partes de la regleta, la numeración sea de menor a

mayor.

• Asegúrese de que las partes de la regleta estén bien unidas y que las figuras

geométricas coincidan, es decir, debe unir (☼ con ☼), (◘ con ◘) y (◙ con ◙).

• Introduzca el tope superior y colóquelo de tal manera que la ventanilla con las

flechas queden del lado de la numeración.


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• Introduzca la columna en la base de plástico asegurándose que esté

totalmente dentro de la ranura de la base y que la numeración quede del lado

izquierdo.

• Indique a la persona que se quite los zapatos.

• Para los casos que tengan pantalones muy largos, realice el doblez hacia

arriba, de tal manera que pueda observar los talones y pies de la persona.

• En caso de peinados altos (chongos, coletas, diademas), indíquele que se los

quite.

• Solicite a la persona que se coloque de espalda a la columna numérica en

posición recta con los brazos a los costados, corrobore que los talones,

pantorrillas, glúteos, espalda y cabeza queden pegados a la columna y los pies

ligeramente separados haciendo un ángulo de 45° gra dos, sin que la persona

se recargue.

• La línea media del cuerpo deberá coincidir con la línea media del estadímetro.

• La cabeza debe estar alineada con respecto al cuerpo, derecha y pegada a la

columna, el punto de referencia que se considera es el vértex (o el punto más

alto del cráneo) y la barbilla debe estar centrada y paralela al suelo.

• Con la mano izquierda tome la barbilla del sujeto a fin de controlar la cabeza y

orientarla hacia el plano de Frankfurt (se refiere a una línea imaginaria que se

marca entre la órbita inferior del ojo y el cartílago prominente del oído medio)

con la mano derecha deslizará el tope móvil hasta tocar la parte coronal de la

cabeza formando un ángulo de 90°.


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Cabeza en posición del Angulo de Frankfurt Posición erguida

• Asegúrese que la posición sea la correcta.

• Después realice la lectura de la talla que está indicada con las flechas que

vienen marcadas en la ventanilla del tope movible del estadímetro; la lectura

deberá tomarse del lado izquierdo de la persona y en forma horizontal para

precisar la lectura.

• Se registrará la talla en el programa de captura, bitácora y tarjeta de

resultados.

Ejemplo: Estatura 165.2=/_1_/_6_/_5_/./_2_/

Centímetros mm

• Los pies marcados deben de quedar centrados en medio de la base del

estadímetro (figura 1)


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Figura 1 posición correcta para medir la talla

Circunferencia de Cintura

� Concepto

La circunferencia de la cintura y cadera son ampliamente utilizados como indicadores

de obesidad abdominal en estudios sobre factores de riesgo vasculares y

metabólicos. También está claro que una gran circunferencia de cintura es el mejor

indicador de grasa intra-abdominal, de grasa visceral y de los factores de riesgo a que

conlleva esto.

� Equipo y Material

- Cinta métrica de fibra de vidrio.

- Lápiz o pluma



�Condiciones generales para la toma de las circunfere ncias.

� Colocar y marcar el punto anatómico de referencia

� Colocar la cinta métrica en circunferencia


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� La cinta no debe de hacer presión y no debe de estar doblada

� La lectura debe de tomarse en centímetros y milímetros

� Técnica de medición

• El adulto deberá estar relajado de frente dejando desnuda la zona en que se

tomara la medida, los brazos cruzados y descansados sobre los hombros, sin

zapatos.

• Se palpa el borde costal inferior y el borde superior de la cresta iliaca de ambos

lados y la ultima costilla flotante de ambos costados para identificar el punto

medio.

• Con la cinta métrica se toma la distancia media axilar y después se hace lo

mismo del lado izquierdo.

• Una vez marcada la media en los 2 lados con un bolígrafo, se coloca la cinta

sin comprimir alrededor de la cintura dejando visible el cero de la cinta para

medir la circunferencia y vigilando que la cinta se encuentre horizontal sin

dobleces, tome la lectura correspondiente. Recuerde que la medición se lee en

centímetros y milímetros.

• Evite que sus dedos queden entre la cinta métrica y el cuerpo del adulto, ya

que esto conduce a tomar una lectura errónea.

• Se realizará 2 veces la medición para que esta sea más precisa y en caso que

tenga duda entre la primera y segunda para corroborar se realizara una

tercera medición.


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Circunferencia de cadera

� Concepto

Es un indicador que evalúa la distribución de tejido adiposo alrededor de la extensión

más grande de las nalgas.



- Lápiz o pluma



� Técnica de medición

• El adulto deberá permanecer de pie con los pies separados unos 20 cm y con

el peso distribuido en forma pareja sobre ambos pies descalzos y con la

menor ropa posible.

• Esta circunferencia se toma horizontalmente en el nivel de máxima extensión

de los glúteos.

• La medida se realiza en la parte más grande o voluminosa de los glúteos.

• La evaluación se puede efectuar en el nivel de los trocánteres.

• El antropometrista adoptara una posición junto al adulto de tal modo que pueda

ver el nivel de extensión máxima de los glúteos y colocar la cinta alrededor de

estos en un plano horizontal.

• La cinta debe estar pegada a la piel pero se debe de procurar no comprimir

demasiado.

• La lectura deberá realizarla del lado izquierdo del individuo, esto para evitar

malos entendidos.


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Medición de la Altura de la Rodilla

� Concepto

Son las maniobras que se realizan para tomar la medición de la distancia entre la

rodilla y el talón en centímetros

Punto tibial lateral (Tíbiale Laterale). Es el punto más proximal y lateral o externo de

la extremidad proximal de la tibia (platillo tibial externo). Con él se pueden determinar

la longitud del muslo, la altura tibial, el perímetro del muslo en su 1/3 medio.



- Lápiz o pluma



-


• Nota: antes de proceder a realizar la medición, se le pedirá al adulto que se

descubra la pierna 3 dedos arriba de la rodilla hasta la altura del muslo, en

caso de que este tenga algún impedimento físico, se le ayudara a realizar esta

maniobra

• Se mide la distancia entre el talón y la parte más alta de la articulación de la

rodilla, por la parte lateral externa, con la pierna flexionada con el adulto

sentado y formando un ángulo de 90° entre el muslo y la pantorrilla

• Colocado el antropometrista delante del adulto, pedirle que flexione la rodilla

formando un ángulo de 90º y/o se sentará para facilitar la localización del

punto.

• Este punto se localiza buscando, en primer lugar, con el dedo pulgar o índice,

la depresión o la interlínea articular de la rodilla, rodeada por tres

protuberancias (epicóndilo femoral, borde anterolateral de la tibia y la cabeza

del peroné); en segundo lugar, y a partir de esta orientación, el antropometrista


23

presiona hacia dentro con la cara lateral del pulgar de la mano derecha hasta

localizar el borde de la tibia y, por último, se palpará hacia atrás hasta localizar

el punto anatómico coincidente con la zona más proximal y externa de la

meseta tibial. Este punto está al menos a un tercio de la distancia entre el

punto anterior y posterior de la rodilla.

• Una vez identificado el punto anatómico, el sujeto se coloca nuevamente en

bipedestación realizándose la marca justo en el lugar en que el dedo o la uña

del pulgar o el índice sienten el borde tibial descrito, comprobándose, como

siempre, que está correctamente señalado.

• Debe medir, de ser posible en la pierna izquierda con el adulto sentado, sin

zapatos y con la rodilla en ángulo recto (en personas encamados se debe

poner la pierna en ángulo de 90°)

• Medir la distancia entre la mano puesta encima de la rodilla y el punto de

contacto del talón con el suelo. Siguiendo una línea recta que debe pasar por

el maléolo externo

• Redondear la medida en 0.5 cm.

• Registrar el resultado.

Altura de la medicion de la rodilla


24

Medición de Altura Sentado

� Concepto

Distancia entre el vértex y el plano de sustentación del entrevistado, medida en cm y

milímetros.


- Fléxometro.

- Escuadra

- Lápiz o pluma




• Es la distancia que existe entre el vértex y los puntos inferiores de la pelvis

(ambos isquion), que se apoyan en el banco.

• Normalmente, esta medida se debería efectuar con el adulto sentado en una

silla y con los pies descalzos y pegados al piso en una superficie plana.

• Se orienta la cabeza del sujeto en el Plano de Frankfort, se le pide que esté

sentado lo más erguido que pueda, tocando con la zona alta de la espalda y

parte posterior de la cabeza en la pared y formando un ángulo de 90º con los

muslos, al igual que la articulación de la rodilla y las manos apoyadas en los

muslos.

• Se registra la medición en su bitácora, recuerde que son centímetros y

milímetros.

• Se le invita a levantarse del banco.

• Este procedimiento se realizará a cada uno de los adultos 2 veces. Si surgiera

alguna duda entre la primera medición y segunda se realiza una tercera para

confirmación.


25

Medidas de Desempeño

La estimación de la capacidad funcional es importante en la evaluación de las

personas en edad avanzada; por lo general se determina esa capacidad en términos

de las actividades cotidianas, como caminar, vestirse y comer. También pueden

efectuarse diversas pruebas funcionales como la fuerza del apretón, el tiempo

necesario para caminar 3 metros y la capacidad para mantenerse en pie sobre una

sola pierna. Estas pruebas son buenos elementos predictivos de la independencia de

la función general y en general se relacionan con la masa corporal y la masa

muscular.

Se piensa que un estado nutricional deficiente y las alteraciones de la composición

del cuerpo se asocian con crecientes problemas del equilibrio y la marcha en las

personas de edad avanzada y, por lo tanto, con el riesgo de sufrir caídas

Antropometrista es importante que las instrucciones para las siguientes pruebas

sean claras y precisas para que el adulto pueda realizarlas, recuerde que de ninguna

manera debe el adulto sentir que está siendo evaluado.


26

Ahora haremos algunos ejercicios para medir su movilidad. Le voy a mostrar cómo

hacer el siguiente ejercicio. Me gustaría que usted tratara de hacerlo. Si cree que no

puede o cree que es peligroso para usted le ruego que me lo diga.

Estando de pié, por favor intente pararse en un solo pié sin apoyarse o agarrarse de

ninguna cosa. Puede intentarlo con cualquiera de sus piernas; después probaremos

con la otra.

Voy a contar el tiempo, así le avisaré cuándo empezar y cuando terminar (DIEZ

SEGUNDOS). Podemos parar en cualquier momento que usted sienta que pierde el

equilibrio.

Comencemos con la pierna con la que se sienta más seguro(a)

Registro de resultados

Se va a registrar el tiempo que el adulto logre permanecer parado en un solo pie tanto

para el pie izquierdo como para el derecho, en caso de que no realice la prueba se

debe de registrar el código que corresponda de acuerdo sea el caso del resultado de

la prueba.


27

Velocidad de la Marcha

Ahora voy a observar como camina usted normalmente. Si usted usa un bastón u otra ayuda para caminar y siente que la necesita para caminar un recorrido corto, puede usarla.

Para esta prueba vamos a buscar un espacio adecuado en donde se pueda realizar este ejercicio.

�� Equipo y Material

- Listón de 3 metros de largo- Cronometro (reloj)- Bitácora de registro

Indicaciones:

Primera Prueba de Velocidad al caminar

Este es nuestro recorrido. Le pediré que usted camine hasta el final del

recorrido a su velocidad acostumbrada, tal como si estuviera caminando en la

calle para ir a la tienda.

Muestre el recorrido al participante.


28

Camine todo el recorrido hasta que pase a la otra orilla de la cinta antes de

parar. Yo caminaré con usted. ¿Usted siente que esto es seguro?

Pida al participante que se ponga de pie con los dos pies tocando la línea de

inicio.

Cuando quiera que comience, yo diré: “Listo, empiece.” Cuando el participante

reconozca esta instrucción diga: “Listo, empiece.”

Presione el botón de empezar para iniciar el cronómetro mientras que el

participante empieza a caminar.

Camine detrás y hacia un lado del participante.

Pare de tomar el tiempo cuando uno de los pies del participante esté

completamente al otro lado de la línea.

3 metros


Se va a registrar el tiempo que el adulto logre pasar uno de los 2 pies completamente

al otro lado la línea de termino del listón, en caso de que no realice la prueba se debe

de registrar el código que corresponda de acuerdo a los códigos de resultado de la


29

pregunta1.18

B. Segunda Prueba de Velocidad al caminar

Ahora le pediré que repita el recorrido. Recuerde que debe caminar a su ritmo

acostumbrado, y siga hasta que pase al otro extremo del recorrido.

Pida al participante que se ponga de pie con los dos pies tocando la línea de

inicio.

Cuando yo quiera que comience, yo diré: “Listo, empiece.” Cuando el

participante reconozca esta instrucción diga: “Listo, empiece.”

Presione el botón de empezar para iniciar el cronómetro mientras que el

participante empieza a caminar.

Camine detrás y hacia un lado del participante.

Pare de tomar el tiempo cuando uno de los pies del participante esté

completamente al otro lado de la línea.

1.17 Tiempo de la Primera

Prueba

Tiempo para recorrer 3 metros

[___|___] . [___|___] Min seg.

Si no pudo realizar la prueba

anote…….00 00

[___][___] . [___][___]

1.18 Si el participante no

intentó o falló en la prueba,

indique el motivo:

Trató, pero no pudo…………………………………..…...1

El participante no pudo mantener

su posición sin ayuda..…………………….…………….2

No intentó, usted se sintió inseguro.................3

No intentó, el participante se sintió inseguro....4

El participante no pudo entender

las instrucciones..………………………………….……..5

Otro (Especifique)_________________.........6

Rehusó hacerla..................................................7

[____]

1.19. Ayudas para el primer

recorrido

Ninguna…........1

Bastón …..........2

Otra…..............7 [____]

> 0 pase a 1.19

Pase

a 1.23


30


Se va a registrar el tiempo que el adulto logre pasar uno de los 2 pies completamente

al otro lado la línea de termino del listón, en caso de que no realice la prueba se debe

de registrar el código que corresponda de acuerdo a los códigos de resultado de la

pregunta 1.21

Prueba de la Fuerza de Presión.

Introducción

La fuerza en las manos afecta las funciones de la vida diaria como el levantar objetos o

cuerpos pesados y normalmente esta fuerza declina con la edad.

Ahora evaluaremos la fuerza de su mano al apretar un objeto. Voy a pedirle que apriete

un objeto tan fuerte como pueda hacerlo, sólo por un par de segundos y luego soltarlo.

Realizaremos la prueba con su mano derecha y su mano izquierda.

�� Equipo y Material

- Dinamómetro


Este equipo se utiliza para medir la fuerza en kilogramos del adulto mayor, realizando

esta prueba en 2 diferentes tiempos, como se muestra en la siguiente imagen:


31

Instrucciones

Le mostraré como hacerlo

• Sugiera al informante que se quite sus anillos u otras joyas similares.

• Usando la mano dominante del informante ajuste el dinamómetro al

entrevistado -de vuelta a la barra para moverla hacia arriba o hacia

abajo- de manera que la barra se apoye en la parte media del dedo

índice y del dedo anular

• En posición de píe, mantenga el dinamómetro en un ángulo de 90° y

apriete el mango durante unos segundos.

• Verifique que el informante esté en la posición correcta: de pie y

formando con el brazo un ángulo de 90°.

• Verifique que el dinamómetro marque cero.

• Explique el procedimiento nuevamente.

• Permita que el informante practique con su mano dominante. Si el

informante no puede usar su mano dominante, practique con la otra

mano y espere 30 segundos entre cada prueba.

• Este procedimiento se deberá realizar 2 veces en cada mano.


32

ENTREVISTADOR:

Verifique la respuesta de la pregunta 1.23. Si la respuesta es código “1” realice 1.26 y

1.27; si la respuesta es código “2” realice 1.27 y si la respuesta es código “3” realice

sólo 1.26

PRIMERA MEDICIÓN SEGUNDA MEDICIÓN

1.26. Haremos dos

mediciones con la

mano izquierda

[____][____].[____] kg

Intentó pero no pudo….. 993.0

No lo intentó………………..999.0

[__][__].[__] kg [____][____].[____] kg

Intentó pero no pudo. 993.0

No lo intentó...………....999.0

[__][__].[__] kg

1.27. Haremos dos

mediciones con la

mano derecha

[____][____].[____] kg

Intentó pero no pudo….. 993.0

No lo intentó…….………..999.0

[__][__].[__]kg [____][____].[____] kg

Intentó pero no pudo. 993.0

No lo intentó...…….…..999.0

[__][__].[__] kg


33

Obtención de Muestras de Sangre Capilar

Sangre capilar : También denominada periférica, es transportada a través de

capilares los cuales son vasos sanguíneos finísimos, interpuestos a través de cuyas

paredes se producen los intercambios entre sangre y tejidos. La piel fría y cianótica es

una fuente de error, esto se evita dando un masaje a la piel hasta que esté rosada y

caliente antes de la punción. La punción se hará preferiblemente en el borde libre del

pulpejo del dedo anular pues resulta más cómodo y accesible. No deberá hacerse en

una parte edematosa o congestionada, ya que es esencial que la sangre pueda fluir

libremente para obtener resultados reproducibles que se puedan comparar con los de

la sangre venosa. Una vez que la punción esté hecha, debe evitarse frotar y exprimir

fuertemente porque también da origen a error.

� Equipo y material General

- Sanitas

- Guantes

- Torunderos

- Torundas impregnadas con alcohol y secas

- Portalancetas (softclix)

- Lancetas

- Contenedor de desechos punzo cortantes

- Bolsas rojas para desechos tóxicos RPBI (torundas y guantes)

- Bolsa negra para deshechos no tóxicos (envoltura de guantes, sanitas, etcétera)


34

Determinación de Hemoglobina Glucosilada

� Concepto

La hemoglobina es una proteína que lleva los glóbulos rojos o hematíes. El azúcar de

la sangre se une a la hemoglobina para formar la hemoglobina A1 (glucosilada).

Si la sangre contiene más azúcar, la hemoglobina glucosilada aumenta y permanece

aumentada durante 120 días. Por esto, la medición de la hemoglobina glucosilada

refleja todas las altas y bajas de glucosa en la sangre en las pasadas 12 semanas.

La hemoglobina A1 es un promedio del nivel de azúcar en los últimos meses,

mientras que un examen para azúcar en la sangre (glucosa) sólo indica el estado de

control de diabetes en un punto determinado.

� Material especifico

- El equipo de A1CNOW consta de:

- A1CNOW Monitor.

- Sobre 1: Kit de dilución de la muestra

- Micropipeta Pasteur

- Recolector capilar

- Criobial con solución

- Porta criobial

- Sobre 2: Cartucho de prueba

- Cartucho


• Coloque el material y el equipo A1CNOW de forma accesible sobre las sanitas,

las cuales estarán sobre una superficie plana y segura.

• Arme el soporte para el críotubo y colóquelo cerca del A1CNOW.

• Tenga a la mano la pipeta para la recolección de la gota.


35

• Quite el capuchón del portalancetas y coloque la lanceta presionando para

fijarla, una vez fija, gire hasta quitar el recubrimiento de la lanceta y coloque la

capucha del portalancetas hasta escuchar un clic.

• En caso que el lugar de punción este frio, pedirle al adulto que sede masajes

en las manos para calentar la zona, esto nos permitirá que obtengamos una

buena gota

• Aseo del sitio de punción. Se realiza la asepsia con una torunda impregnada

con alcohol sobre la cara lateral del pulpejo del dedo anular o medio de

preferencia de la mano que utilice menos, con el fin de quitar la suciedad, el

detritus epitelial y para aumentar la cantidad de sangre en este sitio.

• Deje secar el residuo de alcohol. Cuide que el lugar de punción esté

completamente seco, esperando durante algunos segundos, ya que, de lo

contrario, la sangre no formará una gota redondeada al salir.

• Coloque el portalancetas cargado sobre la cara lateral del dedo y ejerza una

ligera presión sobre la zona elegida para la punción; el disparo de la lanceta

será automático, espere de 2 a 3 segundos antes de retirar el portalancetas.

• Coloque la mano en declive para facilitar la salida de gota de sangre, sin tocar

el dedo y evitando exprimir para no diluir la muestra con líquido de los tejidos y

liberar mas glucosa.

• Esta gota se absorberá colocando la micropipeta de tal manera que la punta

absorba la gota de sangre evitando no tocar la piel a una distancia suficiente

del dedo y cuidando que sea lo suficientemente grande, de manera que se

llene hasta la línea negra y no contenga burbujas de aire para obtener un

resultado óptimo; si esto ocurre, se tendrá que repetir el procedimiento de

recolección.

• Limpie cuidadosamente con una torunda seca el exceso de sangre localizada

alrededor de la pipeta.

• Introduzca la pipeta en el críotubo y de un sólo golpe presione, deje caer la

gota de sangre, saque la pipeta, cierre bien el críotubo y realice de 6 a 8

inversiones vigorosas para diluir la sangre con la sustancia.


36

• Una vez realizada la inversión, coloque el críotubo en la base y destápelo

cuidadosamente.

• Inmediatamente inserte el cartucho en el equipo de A1CNOW.

• Con la segunda pipeta de burbuja, procederá a extraer la sustancia,

inmediatamente traslade la cantidad recolectada al equipo A1CNOW y

presione vigorosamente la pipeta en el círculo de color blanco que viene en el

centro del cartucho sin salirse del él. Recuerde en todo momento que la pipeta

debe estar llena hasta la marca indicada y sin burbujas; si se llega a regar, esto

puede marcar error, por lo que se tendrá que repetir el procedimiento.

• Debe recordar que el equipo cuenta sólo con 10 reactivos, por lo tanto, se

debe tener el cuidado necesario para evitar desperdiciar este material.

Nota : El equipo tiene que mantenerse en temperatura ambiente de 18-28C°. En caso

que la temperatura sea menor, se debe proceder a calentar el cartucho frotándolo o

guardándolo en un lugar caliente para activar el reactivo. Si la temperatura es mayor,

debe conseguirse una hielera y gel congelante para conservar la temperatura.

.

Tome la muestra de sangre capilar. Ponga la muestra en el críotubo.


37

Agite vigorosamente 6-8 veces. Inserte el cartucho y agregue la muestra.


38

Determinación de hemoglobina

� Concepto

Hemoglobina es el componente principal de los glóbulos rojos, del 31% al 34 %, es

una proteína de la sangre que se encarga de transportar el oxígeno desde los

órganos respiratorios hasta los tejidos. Es posible identificar a la hemoglobina como

una heteroproteína ya que es una proteína conjugada (combina una parte proteica

denominada globina con una parte no proteica que se conoce como grupo

prostético).

La técnica de Hemocue es usada ampliamente en estudios de campo, por su facilidad

de hacer las mediciones in situ, sin necesidad de preparar y conservar las muestras.

Es muy reproducible y su precisión u exactitud son muy buenas comparadas con

mediciones de Hb hechas en laboratorio con métodos de citometría de flujo. La

concentración de hemoglobina se obtiene en unidades de gr/dl.

� Material especifico

- Hemocue

- Pilas AA

- Cargador de luz

- Microcubeta reactiva


- Tarjeta de resultados


• Coloque su material y el Hemocue de forma accesible sobre las sanitas, las

cuales estarán sobre una superficie plana y segura.


39

• Muestre al Adulto que el equipo que va a utilizar está limpio y que las lancetas

son nuevas y no han sido utilizadas en ocasiones anteriores.

• Cargar el portalancetas. Destape la punta del portalancetas y coloque una

lanceta presionando para fijarlo, una vez fija, gire hasta quitar el recubrimiento

de la lanceta y coloque nuevamente la parte superior del portalancetas hasta

escuchar un clic.

• Aseo del sitio de punción. Se realiza la asepsia con una torunda impregnada

con alcohol sobre la cara lateral del pulpejo del dedo anular o medio de

cualquiera de las manos, con el fin de quitar la suciedad, el detritus epitelial y

para aumentar la cantidad de sangre en este sitio.

• Deje secar el residuo de alcohol. Cuide que el lugar de punción esté

completamente seco, esperando durante algunos segundos, ya que, de lo

contrario, la sangre no formará una gota redondeada al brotar.

• Técnica de punción. Coloque el portalancetas cargado sobre la cara lateral del

dedo y ejerza una ligera presión sobre la zona elegida para la punción; el

disparo de la lanceta será automático. Enseguida retire el portalancetas.

• La primera gota de sangre. Coloque la mano en declive para facilitar la salida

de una primera gota de sangre, al mismo tiempo que la absorberá para la

prueba de hemoglobina glucosilada, sin tocar el dedo y evitando exprimir o

ordeñar para no diluir la muestra con líquido de los tejidos.

• Coloque la segunda gota de sangre en la microcubeta especial para medir

hemoglobina en el Hemocue. Esta gota se absorberá colocando la cubeta de

tal manera que la punta absorba la gota de sangre, evitando no tocar la piel a

una distancia suficiente del dedo, colocando la zona de color de la cubeta del

Hemocue y cuidando que sea lo suficientemente grande, de manera que se

llene completamente la cubeta y no contenga burbujas de aire para obtener un

resultado óptimo.

• Limpie cuidadosamente con una torunda seca el exceso de sangre localizada

alrededor de la microcubeta.


40

• Introduzca la microcubeta en el Hemocue (encendiendo previamente el botón

que se encuentra en la parte superior izquierda después de la pantalla),

coloque la cubeta llena en el sitio del portacubeta del Hemocue y

posteriormente presione hacia dentro para obtener la medición de

hemoglobina. Para evitar mediciones erróneas, la cubeta debe permanecer en

esta posición mientras no aparezca el resultado en la pantalla lo que tarda de

15-45 segundos.

• Forma de riesgo de resultados de hemoglobina. La cifra de hemoglobina que

aparece en la pantalla digital del Hemocue deberá ser registrada por el

antropometrista en la sección de registro de mediciones biológicas.

• Retirar la cubeta de la ranura y pulsar la tecla para apagar el equipo.

• Desechar la lanceta, torundas utilizadas, guantes y microcubeta que se usó

para la cuantificación de hemoglobina en el recipiente especial destinado para

cada uno de ellos.

� Precauciones para la determinación de hemoglobina

� En el momento de la punción se saca la microcubeta, extraer únicamente una

a la vez.

� Nunca dejar destapado el tubo de Microcubetas de Hemocue.

� Nunca tocar la zona del reactivo de la microcubeta con los dedos.

� Cuando el tubo se ha abierto, se debe poner la fecha ya que tiene 90 días para

usarse después de ser destapado.

� No destapar un tubo nuevo hasta que el anterior se haya terminado por

completo.

� Limpiar semanalmente con una torunda humedecida en alcohol o agua

destilada el área de lectura del equipo, cuidando de no dejar pelusa ni raspar

nada; deje secar todas las partes, introduzca la porta microcubeta

completamente en el aparato.


41

Cuando aparecen las tres rayas fijas en el fotómetro indica que está en proceso de

lectura

Microcubeta obtención de sangre


42

Coloque la cubeta en el soporte para la lectura

El resultado se visualiza después de 15 a 45 segundos


43

Toma de Muestra Sanguínea

Introducción

La determinación del estado de salud de los adultos mayores debe incluir mediciones

de laboratorio, en este estudio se considera la medición de:

Colesterol Total y Colesterol de Lipoproteínas de Alta Densidad (HDLc). El

HDL es el colesterol bueno llamado así porque es factor de "protección" del

sistema cardiovascular.

La Proteína C-reactiva (PCR), es un biomarcador del sistema inmunológico, el

marcador más comúnmente utilizado para medir inflamación e infección.

Además a sido ampliamente utilizado para estudiar la asociación de PCR con

arteriosclerosis, diabetes tipo 2 y enfermedad cardiovascular.

Vitamina D. Además de ser un indicador de la salud ósea, en los últimos años

el déficit de vitamina D se ha vinculado con algunos tipos de cáncer, con la

función muscular y con el equilibrio entre otros.

TSH (Hormona estimulante de tiroides). La forma más común de disfunción

tiroidea en las personas de mayor edad es el hipotiroidismo subclínico. Los

riesgos potenciales del hipotiroidismo subclínico en los ancianos incluyen la

progresión a hipotiroidismo clínico, efectos cardiovasculares, hiperlipidemias, y

efectos neurológicos

Por otra parte, la obtención adecuada de una muestra sanguínea permite realizar

análisis de ella satisfactoriamente, por lo cual es de suma importancia llevar a cabo

este procedimiento lo más seguro, rápido, y fácilmente posible. Es importante, para

garantizar la calidad de la muestra que ésta sea correctamente colectada, que su

origen sea seguro y que los procedimientos de preparación y conservación sean

estrictamente observados (reposo, centrifugado, pipeteo y etiquetado) y que se

conserve adecuadamente hasta su llegada al laboratorio, donde se harán las

determinaciones.


44

El personal que lleva a cabo este procedimiento debe tener presente que el éxito del

trabajo encomendado depende de sus conocimientos, el trato correcto con los sujetos

de estudio y la habilidad para realizar el trabajo.


- Ligadura de látex

- Guantes de látex

- Tubo Vacutainer de tapón rojo

- Aguja Vacutainer

- Aguja Vacutainer verde mariposa

- Aguja Vacutainer azul mariposa

- Sanitas

- Torunderos

- Torundas con alcohol y secas

- Termo individual

- Gradilla

- Gel congelante

- Contenedor de desechos punzo cortantes chico

- Contenedor de desechos punzo cortantes grande

- Bolsa negra de deshechos

- Bolsa roja RPBI

- Etiquetas (Blancas o impresas) para identificación de la muestra

- Marcador indeleble



45

Técnica para Extracción Venosa

• El adulto no necesariamente debe de estar en ayuno la muestra se tomara

casual, es decir a la hora que se encuentre al seleccionado.

• Deberá seleccionar un área adecuada, cómoda y con buena iluminación dentro

del hogar para la toma de muestra.

• Se debe explicar que procedimiento que se le realizará al adulto, para hacerlo

sentir seguro.

• Deberá permanecer sentado de forma que el este cómodo y para la enfermera

sea accesible la toma de muestra.

• se prepara el campo y se acomoda el material necesario para la toma de

muestra, explique que el material es nuevo, completamente estéril y muestre al

adulto la aguja en el momento que la destapa.

• Pídale al adulto que le muestre su antebrazo de ambos brazos, para checar

que vena y brazo son los adecuados para la toma (Ver figura 1), lugar de

donde se recomienda obtener la muestra de sangre.

Figura 1. Venas superficiales del brazo:

1Cefalica, 2 Basílica, 3 Media Basílica, 4 Media Cefálica, 5 Radial accesoria,

6 Cubital superficial y 7 radial superficial


46

• Debe palpar con la punta de los dedos índice y medio la vena para checar el

grosor y la trayectoria de la misma.

• Una vez seleccionado el sitio de punción, aplica un torniquete de 7 a 8 cm por

arriba del pliegue del codo. No se debe de dejar el torniquete por más de 3

minutos (porque esto ocasiona hemolisis) y se recomienda que se le pida al

adulto que cierre el puño para resaltar más las venas.

• Se realiza la asepsia en la zona de punción, debe realizarla del centro a la

periferia y de arriba hacia abajo, rotando su torunda alcoholada. Se deja secar

la zona y no se toca ya.

• Se fija firmemente la vena, con los dedos índice y pulgar distendiendo la piel

del lugar de punción.

• Se realiza la venopunción utilizando el sistema vacutainer y se penetra a

través de la piel con la aguja formando un ángulo de aproximadamente 15° a

30°, con el brazo y con el bisel hacia arriba sigui endo la dirección de la vena. El

tubo se introduce en el dispositivo (la camisa vacutainer), al tener ya la

presencia de sangre en el mismo, se retira el torniquete. Esperar que el tubo se

llene (6ml de sangre total).

NOTA: Este paso es importante ya que necesitamos una buena toma para obtener

suero no hemolizado. Recuerde en todo momento que para realizar una buena toma

debe elegir la aguja de acuerdo al grosor de la vena del adulto seleccionado, además

si acepto la toma de muestra para ADN debe tomar el segundo tubo.


47

• Una vez realizado el procedimiento hay que indicar al adulto que relaje el puño

y se retira el tubo.

• Colocar sobre el sitio de la punción una torunda seca al mismo tiempo que se

retira la aguja con suavidad (la torunda deberá de permanecer entre uno y tres

minutos sobre el sitio de la venopunción).

• Debe rotar de 3 a 5 veces ligeramente la muestra.

• Debe de etiquetar adecuadamente su tubo con el folio que le correspondiente

al adulto seleccionado.

• En todo momento debe estar al pendiente del estado en el que se encuentra el

adulto, esto porque en algunas ocasiones puede marearse al ver la sangre o al

retirar la aguja etc.

�Precauciones Generales.

� El material siempre debe estar a su alcance.

� Debe estar al pendiente del individuo para evitar que se mueva, se pare

o bien se desmaye, ya que estas situaciones traen como consecuencia

accidentes tales como:

� Que se rompa la aguja

� Que se salga la aguja y haya sangrado

� Que se ocasione un hematoma

� Debe pedir al familiar que no haya niños pequeños esto por que en

ocasiones andan corriendo y pueden movernos.

Figura 2. Ejemplo de toma de muestra con sistema Vacutainer®.


48

Procedimiento Después De La Toma De Muestra Sanguínea

• La muestra de sangre total debe conservarse a una temperatura de 2° a 8° C.

• Todas las muestras recolectadas deberán de ser centrifugadas antes de 30

min. y a 2500 RPM durante 15 a 20 minutos.

• Se obtendrá solo el suero del tubo, observar que no se encuentre hemolizado.

• El suero obtenido se separara en 2 alícuotas de 2.00 ml. cada una.

• El coaguló será depositado en el contenedor de punzo cortantes Grande

• Una vez separado y teniendo una cantidad de sueros debe depositarlos al

tanque de Nitrógeno.

• El tanque de Nitrógeno no debe abrirse continuamente puesto ocasiona la fuga

del mismo liquido.

• Cuando el tanque de Nitrógeno está en su capacidad deberá de ser

trasladados al laboratorio del Instituto Nacional de Salud Publica a Cuernavaca

o donde oficina central lo indique.


49

Diagrama de flujo de la antropometría

TOMA 1 DE PRESIÓN ARTERIAL

MEDICION DE ALTURA SENTADO 1 Y 2

MEDICION RODILLA-TALON 1 Y 2

MEDICION PANTORRILLA 1 Y 2

TOMA 2 DE PRESIÓN ARTERIAL

MEDICION DE TALLA 1

MEDICION DE PESO 1

MEDICION DE CINTURA 1

MEDICION DE CADERA 1

MEDICION DE TALLA 2

MEDICION DE PESO 2

MEDICION DE CINTURA 2

MEDICION DE CADERA 2

PRUEBA DE BALANCE

PRUEBA DE MARCHA

PRUEBA DE FUERZA DE PRESIÓN

PRUEBA DE HEMOGLOBINA. GLUCOSILADA

TOMA DE MUESTRA DE SANGRE VENOSA


50

Bibliografía.

Secretaria de Salud. Manual de procedimientos. Toma de medidas clínicas y

antropométricas en el adulto mayor. Subsecretaria de Prevención y Protección para la

Salud. México 2002. Disponible en:

http://www.salud.gob.mx/unidades/cdi/documentos/DOCSAL7518.pdf

Teresa Shamah Levy, Salvador Villalpando Hernández, Juan Rivera Dommarco.

Manual de Procedimientos para proyectos de nutrición. México. Instituto Nacional de

Salud Pública. Diciembre 2006. Disponible en:

http://www.salud.gob.mx/unidades/cdi/documentos/proy_nutricion.pdf

Organización Mundial de la Salud. El estado físico: Uso e Interpretación de la

antropometría. Informe de un Comité de Expertos de la OMS. OMS, Ginebra. 1995.

Disponible en:

http://www.who.int/childgrowth/publications/physical_status_es/en/index.html

Brull DJ, Serrano N, Zito F, Jones L, Montgomery HE, Rumley A, et all. Human CPR gene polymorphism influences CRP levels: Implications for the predictions and pathogenesis of coronary heart disease. Arterioscler Thromb Vasc. Biol, 2003; 23(11); 2063-2069. PMID: 12842840

Cristina Estefanell, Rocío Olivera, et all. Desafíos de la vitamina D. Nuevas propuestas

de suplementación. Arch Pediatr Urug 2010; 81(4): 248-250.

Manual of Procedures Anthropometrics and Biological Sample

51

Apéndice I. Biomarcadores

I. Descripción de la muestra

Se analizaron en total 2.016 muestras distribuidas en cuatro estados. La base de

datos bruta con los resultados de los biomarcadores incluye a los sujetos que

forman parte de la muestra previamente entrevistada por el Instituto Nacional de Estadística Geografía Informática (INEGI), también incluye a 13 sujetos

que participaron únicamente en la toma de muestra de sangre pero no forman

parte de la muestra entrevistada por el INEGI.

El Cuadro 1.1 incluye los identificadores a nivel hogar (CUNICAH), sub-hogar

(SUBHOG_12), e individual (NP) de los sujetos que completaron las muestras

de sangre pero no la entrevista completa con el ENASEM.

Cuadro 1.1. Sujetos que sólo completaron la muestra de sangre

ID del Hogar: CUNICAH (=UNHHID)

ID del Sub-Hogar: SUBHOG_12

ID Individual: NP

3567 1 10 7992 1 20 7995 1 10 8015 1 10 9506 1 20 9513 11 11 10771 11 11 10801 11 10 10943 1 10 13042 0 10 13107 0 10 13110 0 10 14624 0 10


52

Los siguientes resultados descriptivos incluyen la muestra total, incluyendo estos

13 sujetos; sin embargo, estos no se incluyen en el archivo de datos final

disponible en la página web del ENASEM www.MHASweb.org.

El Cuadro 1.2 incluye la distribución de los sujetos según la condición de la

muestra de sangre al momento del análisis de los biomarcadores .

Cuadro 1.2 Distribución de los sujetos de acuerdo a la condición de la muestra de sangre

Tipo de muestra

No parte de la muestra del

ENASEM Muestra del

ENASEM Total

Frecuencia % Frecuencia % Frecuencia % Sin observaciones 11 84.62 1,571 78.43 1,582 78.47

Lipémica 2 15.38 384 19.17 386 19.15 Ligeramente lipémica 0 0.00 1 0.05 1 0.05

Hemolizada 0 0.00 37 1.85 37 1.84 Ligeramente hemolizada 0 0.00 10 0.50 10 0.50

Total 13 100.00 2,003 100.00 2,016 100.00

Se consultó con el INSP sobre la conveniencia de incluir en el análisis las

muestras lipémicas y hemolizadas. La siguiente es su opinión respecto a las

diferencias entre los tipos de muestras:

1. El número de muestras hemolizadas no representa un serio problema, ya

que es relativamente pequeño. Dado que las muestras hemolizadas son la

excepción, y representan un pequeño número dentro de una muestra grande

(menos de 2%), se recomienda considerar incluir el total de la muestra en los

análisis.

http://www.MHASweb.org


53

2. En cuanto al análisis del colesterol total y el HDL, es importante tener en

cuenta que la condición de sueros lipémicos puede ser resultado de una

hiperlipidemia o a que no ayunaron. Así mismo, dado que el colesterol total y el

HDL son biomarcadores interpretables en condiciones de no ayuno, los

resultados de los sueros lipémicos deben ser analizados con precaución.


54

II. Resultados de Colesterol Total (CT)

Para el total de 2,016 muestras analizadas el valor promedio del Colesterol Total

fue de 200,6 mg/dL.

Biomarcador N Media DE Mínimo Máximo CHOLESTEROL_mg_dL 2,016 200.65 46.68 78.00 528.00

Los resultados que a continuación se presentan considera los puntos de corte

que recomienda la NORMA Oficial Mexicana NOM-037-SSA2-20023 para la

prevención, tratamiento y control de las dislipidemias.

Recomendable Limítrofe Alto riesgo TC mg/dL < 200 200-239 ≥ 240

El Cuadro 2.1 muestra la distribución de los resultados de Colesterol Total de

acuerdo a los puntos de corte incluidos anteriormente. Los resultados indican

que 57,6% de la muestra presenta valores recomendables de Colesterol Total,

27% se encuentra dentro del límite y 15,3% tiene valores de alto riesgo.

Cuadro 2.1 Colesterol Total (CT) mg/dL

Frecuencia Porcentage < 200 1,161 57.6 200 a 239 546 27.1 ≥ 240 309 15.3 Total 2,016 100.0

Los resultados incluidos en el Cuadro 2.2 indican que el Estado 2, tiene el mayor

porcentaje de personas con nivel de colesterol de alto riesgo (19,4%).

3http://www.salud.gob.mx/unidades/cdi/nom/037ssa202.html4 El nombre de los 4 estados se eliminó para proteger la identidad de los entrevistados.


55

Cuadro 2.2 Resultados de Colesterol Total por Estado4

Estado < 200 mg/dL 200 to 239 mg/dL ≥ 240 mg/dL Total Fr % Fr % Fr %

Estado 1 201 59.1 98 28.8 41 12.1 340 Estado 2 305 52.4 164 28.2 113 19.4 582 Estado 3 196 61.4 82 25.7 41 12.9 319 Estado 4 459 59.2 202 26.1 114 14.7 775 Total 1,161 57.6 546 27.1 309 15.3 2,016

Los resultados por estado y genero en la Figura 2.1, indican que el porcentaje de

sujetos con niveles de colesterol de alto riesgo es similar entre hombre y

mujeres . En la Figura 2.1, los resultados por estado y de género indican que el

porcentaje de sujetos con niveles de colesterol alto riesgo es similar entre

hombres y mujeres, en 3 estados, a excepción del Estado 3, donde la diferencia

es de 8,1 puntos porcentuales mayor en mujeres que en hombres .

Figura 2.1 Resultados de Colesterol Total por Género y Estado

4 El nombre de los 4 estados se eliminó para proteger la identidad de los entrevistados.

58.0 55.9

68.3

59.6 59.9

50.0 57.0 59.0

0

10

20

30

40

50

60

70

80

Estado 1 Estado 2 Estado 3 Estado 4 Estado 1 Estado 2 Estado 3 Estado 4

Hombres Mujeres

Colesterol <200 mg/dL Colesterol 200-239 mg/dL Colesterol >=240 mg/dL


56

Las estadísticas descriptivas por grupo de edad en el Cuadro 2.3 indican que el

mayor porcentaje de personas dentro de los niveles recomendados de colesterol

se encuentra entre la cohorte más joven. Por otra parte, el porcentaje más bajo

de personas con niveles de colesterol de alto riesgo se encuentra entre la

cohorte más joven (91 %), el porcentaje casi se duplica entre el grupo de 51 a 60

años (17,8 %), y es más alto para la cohorte de más edad de 61 años y mayores

(14,5 %).

Cuadro 2.3 Resultados de Colesterol Total por Grupo de Edad

Colesterol mg/dL

Edad Total ≤ 50 51 a 60 61 y más % % % %

< 200 64.5 51.9 60.7 57.7 200 to 239 26.4 30.3 24.8 27.1 ≥ 240 9.1 17.8 14.5 15.3 Total 100.0 100.0 100.0 100.0


57

III. Resultados Colesterol HDL (C-HDL)

Para el total de 2,016 muestras analizadas el valor promedio del HDL fue de

41,20 mg/dL. A diferencia de los valores anteriores, niveles de HDL altos son

más beneficiosos y niveles bajos son perjudiciales para la salud.

Biomarcador N Media DE Mínimo Máximo HDL mg/dL 2,016 41.20 10.40 17.00 92.00

La NORMA Oficial Mexicana NOM-037-SSA2-2012, para la prevención,

tratamiento y control de las dislipidemias recomienda los siguientes puntos de

corte para la valoración y para el Colesterol de lipoproteínas de alta densidad (C-

HDL).

Recomendable Alto Riesgo HDL mg/dL > 35 ≤ 35

Los resultados que se muestran en el Cuadro 3.1 indican que el Estado 3 tiene

el mayor porcentaje de personas con niveles fuera del rango recomendado para

el HDL (37,0 %).

Cuadro 3.1 Resultados de HDL por Estado5

Estado ≤ 35 mg/dL > 35 mg/dL

Total Fr % Fr %

Estado 1 108 31.8 232 68.2 340 Estado 2 165 28.4 417 71.6 582 Estado 3 118 37.0 201 63.0 319 Estado 4 236 30.5 539 69.5 775 Total 627 31.1 1,389 68.9 2,016



58

La Figura 3.1 indica que el porcentaje de personas con niveles de alto riesgo de

HDL es mayor para los hombres en comparación con las mujeres. En el Estado

1 la diferencia es de 20,9 puntos porcentuales, mientras que en los estados

restantes la diferencia oscila entre 11,9 y 13,6 puntos porcentuales.

Figure 3.1 Resultados de HDL por Género y Estado

El Cuadro 3.2 incluye los resultados de los niveles de HDL por grupos de edad.

Los resultados indican que el mayor porcentaje de las personas con niveles de

alto riesgo de HDL se encuentra entre las personas de 60 años y más (32,5 %).

Cuadro 3.2 Resultados de HDL por Grupo de Edad

HDL mg/dL Edad

Total ≤ 50 51 a 60 60 y más

≤ 35 29.9 29.7 32.5 31.1 > 35 70.1 70.3 67.5 68.9 Total 100.0 100.0 100.0 100.0

44.2 36.4

44.4 37.7

23.3 22.8

32.1 25.8

55.8 63.6

55.6 62.3

76.7 77.2

67.9 74.2

0

10

20

30

40

50

60

70

80

90


Hombres Mujeres

HDL > 35 HDL =< 35


59

IV. Resultados de Proteína C Reactiva (PCR)

Para el total de 2,016 muestras analizadas el valor promedio del PCR fue de

4,25 mg/dL.

Biomarcador N Media DE Mínimo

C-Reactive Protein mg/dL 2,016 4.26 7.20 0.00

La valoración de los resultados de PCR considera los lineamientos de la

American Heart Association6 (Asociación Estadounidense de Cardiología) y los

del laboratorio LAMARKT7, los cuales establecen:

• Se encuentra en bajo riesgo de desarrollar enfermedad cardiovascular si

el nivel de PCR está por debajo de 1,0 mg/dL

• Se encuentra en riesgo promedio de desarrollar enfermedad

cardiovascular si los niveles están entre 1,0 y 3,0 mg/dL

• Se encuentra en alto riesgo de desarrollar enfermedad cardiovascular si el

nivel de PCR está por encima de 3,0 mg/dL

Bajo riesgo Riesgo promedio Alto riesgo

PCR mg/dL < 1.0 1.0-3.0 3.0

Los resultados de PCR por estado indican que el Estado 1 (47,9 %) y el Estado

4 (45 %) tienen los mayores porcentajes de las personas con niveles de PCR de

alto riesgo.

6 http://www.nlm.nih.gov/medlineplus/spanish/ency/article/003356.htm 7 http://www.microelisas.com/pdf/PCR%20us%20CLIA%20%20Monobind.pdf


60

Cuadro 4.1 Resultados de Proteína C Reactiva por Estado8

Estado < 1 mg/dL 1 a 3 mg/dL ≥ 3 mg/dL

Fr % Fr % Fr % Estado 1 67 19.7 148 43.5 125 36.8 Estado 2 107 18.4 196 33.7 279 47.9 Estado 3 79 24.8 124 38.9 116 36.4 Estado 4 143 18.5 283 36.5 349 45.0 Total 396 19.6 751 37.3 869 43.1

La Figura 4.1 muestra que la proporción de mujeres con niveles de alto riesgo de

la PCR es sistemáticamente superior a la proporción de hombres. En el Estado

4, la diferencia es de 16 puntos porcentuales, mientras que en el Estado 3 la

diferencia es de 10,2 puntos porcentuales, 7,8 en el Estado 1, y 5,3 en el Estado

2 .

Figura 4.1 Resultados de Proteína C Reactiva por Género y Estado


28.5

22.2

27.2 26.4

13.8 15.7

23.4

13.3

39.4 38.9 42.4

38.8

46.3

30.0

35.9 35.1 32.1

38.9

30.4 34.9

39.9

54.2

40.6

51.6

0

10

20

30

40

50

60


Hombres Mujeres CRP <1 mg/dL CRP 1 a 3 mg/dL CRP >=3 mg/dL


61

El Cuadro 4.2 incluye los resultados de los niveles de PCR por grupos de edad.

Los resultados indican que los adultos jóvenes tienen el mayor porcentaje de las

personas con niveles de alto riesgo del CRP (48,2 %).

Cuadro 4.2 Resultados de Proteína C Reactiva por Grupo de Edad

PCR mg/dL

Edad Total

≤ 50 51 a 60 60 y más < 1 23.4 17.7 20.5 19.7 1 to 3 28.4 38.3 38.1 37.2 ≥ 3 48.2 44.0 41.4 43.1 Total 100.0 100.0 100.0 100.0


62

V. Resultados de Hormona Estimulante de Tiroides (TSH)

Para el total de 2,0159 muestras analizadas el valor promedio del TSH fue de

2,88 uIU/mL.

Biomarcador N Media DE Mínimo Máximo TSH uIU/mL 2,015 2.88 5.55 0 100

Los siguientes son los valores de referencia para la Hormona Estimulante de

Tiroides (TSH) establecidos en la Guía de Referencia Rápida10 para el

diagnóstico y Tratamiento de hipotiroidismo primario en adultos. Aunque el

diagnóstico de hipotiroidismo (primario, secundario o subclínico) requiere tanto

de la determinación de la hormona estimulante de tiroides como de la Tiroxina

Libre (T4), solo se proporcionan valores de TSH.

Hipotiroidismo secundario

Normal Hipotiroidismo

subclínico Hipotiroidismo

primario

TSH <0.1 0.1-4.49 4.5-10.0 >10 & <40

Los resultados de TSH por estado, en el Cuadro 5.1, indican que el Estado 3

tiene los porcentajes más altos de personas con hipotiroidismo subclínico y

primario, 12,6% y 3,8% respectivamente.

9 Los resultados de TSH no fueron obtenidos para 1 sujeto porque la muestra de sangre no era suficiente. 10

http://www.cenetec.salud.gob.mx/descargas/gpc/CatalogoMaestro/265_IMSS_10_Hipotiroidismo_Primario/GRR_IMSS_262_10.pdf


63

Cuadro 5.1 Resultados de TSH por Estado11

Estado <0.1 uIU/mL 0.1-4.49

uIU/mL 4.5-1.0 uIU/mL

>10 & <40 mg/dL Total

Fr % Fr % Fr % Fr % Estado 1 2 0.6% 295 86.8% 33 9.7% 10 2.9% 340 Estado 2 9 1.5% 522 89.7% 39 6.7% 12 2.1% 582 Estado 3 2 0.6% 264 83.0% 40 12.6% 12 3.8% 318 Estado 4 5 0.6% 687 88.6% 70 9.0% 13 1.7% 775 Total 18 0.9% 1,768 87.7% 182 9.0% 47 2.3% 2,015

Los resultados por estado y género, en la Figura 5.1, indican que el porcentaje

de sujetos con hipotiroidismo subclínico y primario es mayor para las mujeres

comparado con los hombres, a excepción del Estado 1.

Figure 5.1 Resultados de TSH por Género y Estado


0.0 1.0 1.3 1.7 0.0 1.0 0.3 0.8

85.5 87.6 91.5

22.0

84.9 81.8

91.1 87.1

14.5 11.4

7.2 9.8 15.1 17.2

8.6 12.1

0

10

20

30

40

50

60

70

80

90

100


Hombres Mujeres

CRP <.1 uIU/mL CRP .1-4.49 uIU/mL CRP >=4.5 uIU/mL


64

El Cuadro 5.2 incluye los resultados de los niveles de TSH por grupos de edad.

Los resultados indican que el grupo de adultos jóvenes tienen el mayor

porcentaje de personas con nivel de TSH normal: 89,80% para los adultos de 50

años o menores y 89.22 para los adultos de 51 a 60 años de edad, en

comparación con 86,17 para los adultos de 60 años o más.

Cuadro 5.2 Resultados de TSH por Grupo de Edad

TSH uIU/mL Edad

Total ≤ 50 51 a 60 60 y más

<0.1 1.02 0.90 0.87 0.90 .1 to 4.49 89.80 89.22 86.17 87.71 4.5 to 1.0 7.14 7.83 10.35 9.06 >10 & <40 2.04 2.05 2.62 2.61 Total 100.00 100.00 100.00 100.00


65

VI. Resultados de Vitamina D

Para el total de 2,01312 muestras analizadas el valor promedio del TSH fue de

24,21 ng/dL.

Biomarcador N Media DE Mínimo Máximo Vitamina D ng/dL 2,016 24.21 8.62 4.7 92.3

Los siguientes son los puntos de corte para Vitamina D, parte de las directrices

para la evaluación de la deficiencia de Vitamina D.

Deficiencia Normal Vitamina D ng/dL <20 ≥20

Los resultados en el Cuadro 6.1 indican que el Estado 1 tiene el mayor

porcentaje de personas con deficiencia de vitamina D (54,7%), seguido por el

Estado 2 (36,8%).

Cuadro 6.1 Resultados de Vitamina D por Estado13

Estado < 20 ng/dL ≥ 20 ng/dL

Total Fr % Fr %

Estado 1 186 54.7% 154 45.3% 340 Estado 2 214 36.8% 368 63.2% 582 Estado 3 95 30.0% 222 70.0% 317 Estado 4 154 19.9% 620 80.1% 774 Total 649 32.2% 1,36

467.8% 2,013

La Figura 6.1 indica que el porcentaje de personas con deficiencia de vitamina D

es consistentemente más bajo para los hombres en comparación con las

12 Los resultados de Vitamina D no fueron obtenidos para 3 sujetos porque la muestra de sangre no era suficiente. 13 El nombre de los 4 estados se eliminó para proteger la identidad de los entrevistados.


66

mujeres; la diferencia oscila entre 4,4 (en el Estado 4) y 25,7 puntos

porcentuales (en el Estado 3).

Figura 6.1 Resultados de Vitamina D por Género y Estado

El Cuadro 6.2 incluye los resultados de los niveles de Vitamina D por grupos de

edad. Los resultados indican que el mayor porcentaje de personas con

deficiencia de vitamina D se encuentra entre las personas de 60 años y más

(38,0%).

Cuadro 6.2 Resultados de Vitamina D por Grupo de Edad

Vitamina D ng/dL

Edad Total ≤ 50 51 a 60 60 y más

< 20 27.5 25.8 38.0 32.2 ≥ 20 72.5 74.2 62.0 67.8 Total 100.0 100.0 100.0 100.0

46.4

27.1

14.4 17.2

60.4

43.3 40.1

21.6

53.6

72.9

85.6 82.8

39.6

56.6

81.8 78.4

0

10

20

30

40

50

60

70

80

90


Hombres Mujeres

Vitamina D <20 ng/mL Vitamina D >=20 ng/mL


67

Apéndice II. Nota Técnica sobre el Método de Análisis de Laboratorio para la Determinación de Colesterol

Se recibieron un número de muestras de suero que no fueron tomadas en ayuno

y algunas con evidencia de grasa en la muestra (lipémicas). De acuerdo con las

normas del equipo Architect usado para las determinaciones, las muestras

fueron centrifugadas, con lo cual se formó una capa de grasa en la parte

superior del tubo (ver figura 1 anexa), de dimensiones variables y que parte fue

absorbida por la pipeta del brazo del robot del equipo, con lo cual las mediciones

se vieron afectadas.

Figura 1. Efecto de la Centrifugación sobre los Lípidos y el Pipeteo del Robot

Adicionalmente, a petición de la Dra. Mara Téllez-Rojo se centrifugaron 12

muestras que mostraban niveles muy altos de colesterol. Una vez que se

revisaron estos resultados, se analizó el procedimiento utilizado para la medición


68

inicial. Una vez hecho este análisis se decidió repetir todas el análisis de todas

las muestras “sin centrifugación” para que no apareciera la capa de grasa y se

distribuyera en la muestra. Se repitió análisis de las 12 muestras de sangres, se

volvieron a medir y se compararon con un segundo método más drástico que

consiste en calentar la muestra a 38°C durante 5 min para asegurar que la grasa

se disuelva y no presente mayor trabajo para medirla. Este método no se usa

comúnmente en las muestras de suero, ya que afecta otras substancias las

cuales podrían ser medidas más tarde.

La línea azul en la Figura 2 representa las mediciones hechas después de la

centrifugación, las cuales muestran resultados más altos y menos variabilidad en

comparación con las otras tres líneas. Las línea roja y verde representan las

mediciones hechas sin centrifugación; y, finalmente, la línea azul con la 'x'

representa las mediciones realizadas después de calentar la muestra. Como se

puede observar en la figura, las mediciones que no fueron centrifugadas son

altamente comparables y tienen mayor variabilidad en contraste con la línea que

representa las mediciones después de la centrifugación


69

Figura 2. Análisis de 12 Muestras (enviadas para verificación por la Dra. Téllez)

Para asegurar la calidad de las mediciones, se midieron de manera simultánea

sueros de control de calidad, en cantidad doble de lo habitual. Las muestras de

control de calidad se incluyeron al principio del análisis, y se intercalaron cada

50 muestras y al final del análisis. Como se indica en las mediciones que se

presentan en un gráfico de Levy-Jennings a continuación, los resultados

entregados son de buena calidad. Se utilizaron los criterios de Westgard para

interpretar los resultados de control de calidad, los cuales establecen que el

medio determinado por la fabricación no debe exceder de ± 1 desviación

CO

LES

TER

OL m

g/dL


70

estándar (DE). Las Figuras 3 y 4 muestran los resultados de suero sanguíneo de

acuerdo con el criterio establecido por las directrices en Merck Colesterol 2015;

la primera figura presenta una concentración media de 101 mg / dL y la segunda

presenta una concentración media de 242 mg / dL. Los resultados indican que la

diferencia en las mediciones de control de calidad no exceden mas de ± 1 SD.

Figura 3. Curvas de Levy-Jennings. Suero control de calidad Merck Colesterol #2015, lote 14431, con concentración teórica de 101 mg/dL.; corridas en varias mediciones entre el 5 y el 16 de Febrero 2015.


71

Figure 4. Curvas de Levy-Jennings. Suero control de calidad Merck Colesterol #2015, lote 14431, con concentración teórica de 242 mg/dL.; corridas en varias mediciones entre el 5 y el 16 de Febrero 2015.


72

Conclusiones

Se analizaron de nuevo todas las 2.016 muestras sin el método de

centrifugación. Los resultados ‘corregidos’ que se presentan aquí, son buenas

mediciones según lo confirmado por los resultados de control de calidad que se

presentaron anteriormente. Sin embargo, estas muestras no fueron

originalmente tomadas en ayuno, lo cual no cumple con la recomendación de la

ATP II, que recomienda un mínimo de 12 horas de ayuno y el punto de corte

internacional >200 mg/dL para diagnóstico de hipercolesterolemia.

Finalmente, en la siguiente tabla se comparan los resultados de prevalencia de

hipercolesterolemia con los de ENSANUT 2012 y los de la ENASEM.Como se

puede ver, los resultados indican que la prevalencia de la hipercolesterolemia es

similar en ambos estudios.

Tabla 1. Comparación de Prevalencias de Colesterol, ENASEM y ENSANUT 2012

Edad Colesterol

> 200 mg/dL (%)ENASEM 2012 ENSANut 2012

<30 23.81 19.7 31-40 36.65 27.9 41-50 39.56 37.9 51-60 34.12 36.7 61-70 45.32 42.1 71-80 38.26 56.8 >81 34.86 33.2


73

Apéndice III. Material de Instrucciones de Architect®

En la siguiente tabla se enumeran los contenidos de los materiales incluidos en el Apéndice III. Cada paquete contiene la información utilizada por el laboratorio para realizar los ensayos.

Colesterol …………………………………

Ultra HDL (UHDL) ………………………… Page 81

CRP Vario …………………………………. Page 88

TSH …………………………………………. Page 97

TSH Controles ……………………………… Page 104

TSH Calibradores ….………………………. Page 105

25-OH Vitamin D …………………………… Page 106

1

CHOLESTEROL

This package insert contains information to run the Cholesterol assay on the ARCHITECT c Systems™ and the AEROSET System.

NOTE: Changes Highlighted

NOTE: This package insert must be read carefully prior to product use. Package insert instructions must be followed accordingly. Reliability of assay results cannot be guaranteed if there are any deviations from the instructions in this package insert.

Customer SupportUnited States: 1-877-4ABBOTTCanada: 1-800-387-8378 (English speaking customers)

1-800-465-2675 (French speaking customers)International: Call your local Abbott representative

Symbols in Product Labeling

Calibrators 1 and 2 Catalog number/List number

Concentration Serial number

Authorized Representative in the European Community

Consult instructions for use

Ingredients Manufacturer

In vitro diagnostic medical device Temperature limitation

Batch code/Lot number Use by/Expiration date

Reagent 1

CHOLESTEROL 7D62-20

30-3126/R3

ABBOTT LABORATORIESAbbott Park, IL 60064, USA

ABBOTTMax-Planck-Ring 265205 WiesbadenGermany+49-6122-580

September 2006©2002, 2006 Abbott Laboratories

2

NAMECHOLESTEROL

INTENDED USEThe Cholesterol assay is used for the quantitation of cholesterol in human serum or plasma.

SUMMARY AND EXPLANATION OF TESTMeasurement of serum cholesterol levels can serve as an indicator of liver function, biliary function, intestinal absorption, propensity toward coronary artery disease, and thyroid function. Cholesterol levels are important in the diagnosis and classification of hyperlipoproteinemias. Stress, age, gender, hormonal balance, and pregnancy affect normal cholesterol levels.1

The Adult Treatment Panel of the National Cholesterol Education Program (NCEP) recommends that all adults 20 years of age and over should have a fasting lipoprotein profile (total cholesterol, LDL cholesterol, HDL cholesterol, and triglyceride) once every five years to screen for coronary heart disease risk.2

PRINCIPLES OF PROCEDUREThe use of enzymes to assay cholesterol has been studied by many investigators.3,4 This reagent is based on the formulation of Allain, et al.5 and the modification of Roeschlau6 with further improvements to render the reagent stable in solution.Cholesterol esters are enzymatically hydrolyzed by cholesterol esterase to cholesterol and free fatty acids. Free cholesterol, including that originally present, is then oxidized by cholesterol oxidase to cholest-4-ene-3-one and hydrogen peroxide. The hydrogen peroxide combines with hydroxybenzoic acid (HBA) and 4-aminoantipyrine to form a chromophore (quinoneimine dye) which is quantitated at 500 nm.

Methodology: Enzymatic

REAGENTS

Reagent Kit 7D62 Cholesterol is supplied as a liquid, ready-to-use, single

reagent kit which contains:

10 x 84 mL

Estimated tests per kit: 3,032Calculation is based on the minimum reagent fill volume per kit.

Reactive Ingredients ConcentrationCholesterol Oxidase (Microbial) > 200 U/LCholesterol Esterase (Microbial) > 500 U/LPeroxidase (Horseradish) > 300 U/L4-Aminoantipyrine 0.25 mmol/LHBA 10 mmol/L

The Abbott Clinical Chemistry Cholesterol reagent is certified to be traceable to the National Reference System for Cholesterol, against the Abell-Kendall reference method in a CDC-Certified Cholesterol Reference Method Laboratory Network (CRMLN).

REAGENT HANDLING AND STORAGE

Reagent HandlingRemove air bubbles, if present in the reagent cartridge, with a new applicator stick. Alternatively, allow the reagent to sit at the appropriate storage temperature to allow the bubbles to dissipate. To minimize volume depletion, do not use a transfer pipette to remove the bubbles.

CAUTION: Reagent bubbles may interfere with proper detection of reagent level in the cartridge, causing insufficient reagent aspiration which could impact results.

Reagent StorageUnopened reagents are stable until the expiration date when stored at 2 to 8°C.Reagent stability is 30 days if the reagent is uncapped and onboard.

WARNINGS AND PRECAUTIONS

Precautions for Users1. For in vitro diagnostic use.2. Do not use components beyond the expiration date.3. Do not mix materials from different kit lot numbers.4. Contains nonsterile bovine serum albumin.5. CAUTION: This product requires the handling of human specimens.

It is recommended that all human sourced materials are consideredpotentially infectious and be handled in accordance with the OSHAStandard on Bloodborne Pathogens.7 Biosafety Level 28 or otherappropriate biosafety practices9,10 should be used for materials thatcontain or are suspected of containing infectious agents.

SPECIMEN COLLECTION AND HANDLING

Suitable SpecimensSerum and plasma are acceptable specimens. The National Cholesterol Education Program (NCEP) recommends using fasting specimens.2

• Serum: Use serum collected by standard venipuncture techniquesinto glass or plastic tubes with or without gel barriers. Ensurecomplete clot formation has taken place prior to centrifugation.Separate serum from red blood cells or gel as soon after collectionas possible.

Some specimens, especially those from patients receivinganticoagulant or thrombolytic therapy, may take longer to completetheir clotting processes. Fibrin clots may subsequently form in thesesera and the clots could cause erroneous test results.

• Plasma: Use plasma collected by standard venipuncture techniquesinto glass or plastic tubes. Acceptable anticoagulants are lithiumheparin (with or without gel barrier) and sodium heparin. Ensurecentrifugation is adequate to remove platelets. Separate plasma fromred blood cells or gel as soon after collection as possible.

Refer to the specimen collection tube manufacturer’s instructions for processing and handling requirements.

For total sample volume requirements, refer to the instrument-specific ASSAY PARAMETERS section of this package insert and Section 5 of the instrument-specific operations manual.

Specimen Storage

Serum and plasma

Temperature MaximumStorage

Bibliographic Reference

20 to 25°C 7 days 112 to 8°C 7 days 11, 12-20°C 3 months 11

Guder et al.11 suggest storage of frozen specimens at -20°C for no longer than the time interval cited above. However, limitations of laboratory equipment make it necessary in practice for clinical laboratories to establish a range around -20°C for specimen storage. This temperature range may be established from either the freezer manufacturer’s specifications or your laboratory standard operating procedure(s) for specimen storage.

NOTE: Stored specimens must be inspected for particulates. If present, mix and centrifuge the specimen to remove particulates prior to testing.

PROCEDURE

Materials Provided 7D62 Cholesterol Reagent Kit

Materials Required but not Provided• 1E65 Multiconstituent Calibrator, 3 x 5 mL• Control Material• Saline (0.85% to 0.90% NaCl) for specimens that require dilution

3

PROCEDURE (Continued)

Assay ProcedureFor a detailed description of how to run an assay, refer to Section 5 of the instrument-specific operations manual.

Specimen Dilution ProceduresThe ARCHITECT c Systems and the AEROSET System have automatic dilution features; refer to Section 2 of the instrument-specific operations manual for additional information.

Serum and plasma: Specimens with cholesterol values exceeding 705 mg/dL (18.26 mmol/L) are flagged and may be diluted using the Automated Dilution Protocol or the Manual Dilution Procedure.

Automated Dilution ProtocolIf using the Automated Dilution Protocol, the system performs a 1:4 dilution of the specimen and automatically corrects the concentration by multiplying the result by the appropriate dilution factor.

Manual Dilution ProcedureManual dilutions should be performed as follows:

• Use saline (0.85% to 0.90% NaCl) to dilute the sample.

• The operator must enter the dilution factor in the patient or controlorder screen. The system uses this dilution factor to automatically correct the concentration by multiplying the result by the entered factor.

• If the operator does not enter the dilution factor, the result mustbe multiplied by the appropriate dilution factor before reporting the result.

NOTE: If a diluted sample result is flagged indicating it is less than the linear low limit, do not report the result. Rerun using an appropriate dilution.

For detailed information on ordering dilutions, refer to Section 5 of the instrument-specific operations manual.

CALIBRATIONCalibration is stable for approximately 30 days (720 hours) and is required with each change in reagent lot number. Verify calibration curve with at least two levels of controls according to the established quality control requirements for your laboratory. If control results fall outside acceptable ranges, recalibration may be necessary.For a detailed description of how to calibrate an assay, refer to Section 6 of the instrument-specific operations manual.For information on calibrator standardization, refer to the Multiconstituent Calibrator package insert.

QUALITY CONTROLThe following is the recommendation of Abbott Laboratories for quality control. As appropriate, refer to your laboratory standard operating procedure(s) and/or quality assurance plan for additional quality control requirements and potential corrective actions.

• Two levels of controls (normal and abnormal) are to be run every24 hours.

• If more frequent control monitoring is required, follow the establishedquality control procedures for your laboratory.

• If quality control results do not meet the acceptance criteriadefined by your laboratory, patient values may be suspect. Followthe established quality control procedures for your laboratory.Recalibration may be necessary.

• Review quality control results and acceptance criteria following achange of reagent or calibrator lot.

RESULTSRefer to the instrument-specific operations manual for information on results calculations.

• ARCHITECT System Operations Manual—Appendix C• AEROSET System Operations Manual—Appendix ARepresentative performance data are given in the EXPECTED VALUES and SPECIFIC PERFORMANCE CHARACTERISTICS sections of this package insert. Results obtained in individual laboratories may vary.

LIMITATIONS OF THE PROCEDURERefer to the SPECIMEN COLLECTION AND HANDLING and SPECIFIC PERFORMANCE CHARACTERISTICS sections of this package insert.

EXPECTED VALUES

Reference Range

Serum/Plasma

Range (mg/dL) Range (mmol/L)

Child13

Desirable < 170 < 4.40Borderline 170 to 199 4.40 to 5.15High ≥ 200 ≥ 5.18

Adult2

Desirable < 200 < 5.18Borderline 200 to 239 5.18 to 6.19High ≥ 240 ≥ 6.22

To convert results from mg/dL to mmol/L, multiply mg/dL by 0.0259.The National Cholesterol Education Program (NCEP) Adult Treatment Panel III Report2 recommends the adult classification shown above. Laboratories should follow recommendations for lipid ranges effective in their locale if they differ from those of the NCEP.

SPECIFIC PERFORMANCE CHARACTERISTICS

LinearityCholesterol is linear up to 705 mg/dL (18.26 mmol/L). Linearity was verified using Clinical and Laboratory Standards Institute (CLSI) protocol NCCLS EP6-P.14

Limit of Detection (LOD)The LOD for Cholesterol is 5.0 mg/dL (0.13 mmol/L). The LOD is the mean concentration of an analyte-free sample + 2 SD, where SD = the pooled, within-run standard deviation of the analyte-free sample. A study performed on an ARCHITECT c System and an AEROSET System produced an LOD for the Cholesterol assay of 0.80 mg/dL (0.021 mmol/L).

Limit of Quantitation (LOQ)The LOQ for Cholesterol is 6.2 mg/dL (0.161 mmol/L). The LOQ is the analyte concentration at which the CV = 20%.

Interfering Substances15

Interference studies were conducted using CLSI protocol NCCLS EP7-P.16 Interference effects were assessed by Dose Response and Paired Difference methods, at the medical decision level of the analyte.

InterferingSubstance

Interferent Concentration

N Target (mg/dL)

Observed (% of Target)

Bilirubin7.5 mg/dL (128 µmol/L) 4 252.3 91.715 mg/dL (257 µmol/L) 4 252.3 86.8

Hemoglobin750 mg/dL (7.5 g/L) 4 241.1 109.5

1,000 mg/dL (10.0 g/L) 4 241.1 111.9

Intralipid1,000 mg/dL (10.0 g/L) 4 236.1 102.52,000 mg/dL (20.0 g/L) 4 236.1 101.9

Ascorbate1.5 mg/dL (85 µmol/L) 4 282.2 98.7

3 mg/dL (170 µmol/L) 4 282.2 97.6

Bilirubin solutions at the above concentrations were prepared by addition of a bilirubin stock to human serum pools. Hemoglobin solutions at the above concentrations were prepared by addition of hemolysate to human serum pools. Intralipid solutions at the above concentrations were prepared by addition of Intralipid to human serum pools. Ascorbate solutions at the above concentrations were prepared by addition of ascorbic acid to human serum pools.

PrecisionThe imprecision of the Cholesterol assay is ≤ 3% Total CV. Representative data from studies using CLSI protocol NCCLS EP5-A17 are summarized below.

Control Level 1 Level 2N 80 80

Mean (mg/dL) 261.4 129.2

Within RunSD 1.98 0.78

%CV 0.8 0.6

Between RunSD 1.01 1.03

%CV 0.4 0.8

Between DaySD 3.36 1.64

%CV 1.3 1.3

TotalSD 4.03 2.09

%CV 1.5 1.6

4

SPECIFIC PERFORMANCE CHARACTERISTICS (Continued)

Method ComparisonCorrelation studies were performed using CLSI protocol NCCLS EP9-A.18

Serum results from the Cholesterol assay on the AEROSET System were compared with those from a commercially available enzymatic methodology.

Serum results from the Cholesterol assay on the ARCHITECT c System were compared with the Cholesterol assay on the AEROSET System.

AEROSETvs. Comparative

Method

ARCHITECTvs. AEROSET

N 79 101

Y - Intercept 0.933 -0.840

Correlation Coefficient 0.993 0.993

Slope 1.016 0.979

Range (mg/dL)* 70.6 to 416.8 39.5 to 687.6

*AEROSET Range

BIBLIOGRAPHY1. Burtis CA, Ashwood ER, editors. Tietz Fundamentals of Clinical

Chemistry, 5th ed. Philadelphia, PA: WB Saunders; 2001:480–5.

2. Executive summary of the third report of the National CholesterolEducation Program (NCEP) Expert Panel on detection, evaluation,and treatment of high blood cholesterol in adults (Adult TreatmentPanel III). JAMA 2001;285:2486–97.

3. Flegg HM. An investigation of the determination of serumcholesterol by an enzymatic method. Ann Clin Biochem 1973;10:79–84.

4. Richmond W. Preparation and properties of a cholesterol oxidasefrom Nocardia sp. and its application to the enzymatic assay of totalcholesterol in serum. Clin Chem 1973;19(12):1350–6.

5. Allain CC, Poon LS, Chan CS, et al. Enzymatic determination oftotal serum cholesterol. Clin Chem 1974;20(4):470–5.

6. Roeschlau P, Bernt E, Gruber WA. Enzymatic determination of totalcholesterol in serum. Z Klin Chem Klin Biochem 1974;12:226.

7. US Department of Labor, Occupational Safety and HealthAdministration. 29 CFR Part 1910.1030, Occupational Exposure toBloodborne Pathogens.

8. US Department of Health and Human Services. Biosafety inMicrobiological and Biomedical Laboratories. HHS Publication(CDC), 4th ed. Washington, DC: US Government Printing Office,May 1999.

9. World Health Organization. Laboratory Biosafety Manual. Geneva:World Health Organization, 2004.

10. Sewell DL, Bove KE, Callihan DR, et al. Protection of LaboratoryWorkers from Occupationally Acquired Infections; ApprovedGuideline—Third Edition (M29-A3). Wayne, PA: Clinical andLaboratory Standards Institute, 2005.

11. Guder WG, Narayanan S, Wisser H, et al. List of analytes—preanalytical variables. Annex In: Samples: From the Patient to theLaboratory. Darmstadt, Germany: GIT Verlag; 1996:Annex 12–3.

12. US Pharmacopeial Convention, Inc. General notices. In: USPharmacopeia National Formulary, 1995 ed (USP 23/NF 18).Rockville, MD: The US Pharmacopeial Convention, Inc; 1994:11.

13. American Academy of Pediatrics, Committee on Nutrition.Cholesterol in childhood. Pediatrics 1998:101(1);141–7.

14. Passey RB, Bee DE, Caffo A, et al. Evaluation of the Linearityof Quantitative Analytical Methods; Proposed Guideline (EP6-P).Villanova, PA: The National Committee for Clinical LaboratoryStandards, 1986.

15. Young DS. Effects of Drugs on Clinical Laboratory Tests, 4th ed.Washington, DC: AACC Press; 1995:3-143–3-163.

16. Powers DM, Boyd JC, Glick MR, et al. Interference Testing inClinical Chemistry; Proposed Guideline (EP7-P). Villanova, PA: TheNational Committee for Clinical Laboratory Standards, 1986.

17. Kennedy JW, Carey RN, Coolen RB, et al. Evaluation of PrecisionPerformance of Clinical Chemistry Devices; Approved Guideline(EP5-A). Wayne, PA: The National Committee for ClinicalLaboratory Standards, 1999.

18. Kennedy JW, Carey RN, Coolen RB, et al. Method Comparison andBias Estimation Using Patient Samples; Approved Guideline (EP9-A). Wayne, PA: The National Committee for Clinical LaboratoryStandards, 1995.

TRADEMARKSAEROSET and ARCHITECT are registered trademarks of Abbott Laboratories.

c System is a trademark of Abbott Laboratories.

All other trademarks, brands, product names, and trade names are the property of their respective companies.

5

Cholesterol Serum/Plasma—Conventional and SI Units

Cholesterol Serum/Plasma—Conventional Units

Cholesterol Serum/Plasma—SI Units

† The linear low value (Low-Linearity) is LOQ rounded up to the number of decimal places defined in the decimal places parameter field.‡ Refer to concentration specified on calibrator labeling or value sheet.†† Displays the number of decimal places defined in the decimal places parameter field.

ARCHITECT c SYSTEMS ASSAY PARAMETERS

Configure assay parameters — General

● General о Calibration о SmartWash о Results о InterpretationAssay: Chol Type: Photometric Version: 1

Number: 1018● Reaction definition о Reagent / Sample о Validity checksReaction mode: End up

Primary Secondary Read timesWavelength: 500 / 660 Main: 31 – 33

Last required read: 33Absorbance range: ___ – ___ Color correction: ___ – ___Sample blank type: None

о Reaction definition ● Reagent / Sample о Validity checksR1

Reagent: CHOL0 Reagent volume: 240Diluent: Saline Water volume: ___

Diluent dispense mode:Type 0 Dispense mode: Type 0

Diluted DefaultDilution name Sample sample Diluent Water Dilution factor dilutionSTANDARD : 2.4 ___ ___ ___ = 1:1.00 ●1:4 : 25.0 2.4 75 ___ = 1:4.00 о

_________ : ___ ___ ___ ___ = о

о Reaction definition о Reagent / Sample ● Validity checksReaction check: None

Maximum absorbance variation: ___

Configure assay parameters — Calibration

о General ● Calibration о SmartWash о Results о InterpretationAssay: Chol Calibration method: Linear

● Calibrators о Volumes о Intervals о Validity checksCalibrator set: Calibrator level: Concentration:

MCC Blank: Water 0††

Cal 1: MCC1 ‡Replicates: 3 [Range 1 – 3] Cal 2: MCC2 ‡

о Calibrators ● Volumes о Intervals о Validity checksCalibrator: MCC Diluted

Calibrator level Sample sample Diluent WaterBlank: Water 2.4 ___ ___ ___Cal 1: MCC1 2.4 ___ ___ ___Cal 2: MCC2 2.4 ___ ___ ___

о Calibrators о Volumes ● Intervals о Validity checksCalibration intervals:

Full interval: 720 (hours)Calibration type:

Adjust type: None

о Calibrators о Volumes о Intervals ● Validity checksBlank absorbance range: _____ – _____

Span: Blank – BlankSpan absorbance range: _____ – _____

Expected cal factor: 0.00Expected cal factor tolerance %: 0

Configure assay parameters — SmartWash

о General о Calibration ● SmartWash о Results о InterpretationAssay: Chol

COMPONENT REAGENT / ASSAY WASH Volume ReplicatesR1 ALBP0 Water 300 1

Cuvette Trig 10% Detergent B *** 345

*** Select “Detergent B” for software prior to version 2.2.

Configure assay parameters — Results — Conventional Units

о General о Calibration о SmartWash ● Results о InterpretationAssay: Chol Result units: mg/dL

Assay defaults:Low-Linearity: 7†

High-Linearity: 705Gender and age specific ranges:GENDER AGE (UNITS) NORMAL EXTREMEEither 0 – 130 (Y) 0 – 199

Configure result units — Conventional UnitsAssay: Chol

Version: 1Result units: mg/dL

Decimal places: 0 [Range 0 – 4]Correlation factor: 1.0000

Intercept: 0.0000

Configure assay parameters — Results — SI Units

о General о Calibration о SmartWash ● Results о InterpretationAssay: Chol Result units: mmol/L

Assay defaults:Low-Linearity: 0.17†

High-Linearity: 18.26Gender and age specific ranges:GENDER AGE (UNITS) NORMAL EXTREMEEither 0 – 130 (Y) 0.00 – 5.17

Configure result units — SI UnitsAssay: Chol

Version: 1Result units: mmol/L


Intercept: 0.0000

6

Cholesterol Serum/Plasma—Conventional Units Cholesterol Serum/Plasma—SI Units

AEROSET SYSTEM ASSAY PARAMETERS

Refer to Assay Configuration in Section 2 of the AEROSET System Operations Manual for information regarding assay parameters.* User defined or instrument defined.** The linear low value (L-Linear Range) is LOQ rounded up to the number of decimal places defined in the decimal places parameter field.

Assay Configuration: Outline PageAssay Name Assay # LineChol 18 B-LineQuantitative RangesMin Text Min Panic-L L-Reference-H Panic-H Max Max Text

* 0.0* 0.0 0 199 0.0 0.0* *

7** L-Linear Range-H 705

Reference Ranges*Age Male Female

0 Year0 Year0 Year

0.0 – 0.0 0.0 – 0.00.0 – 0.0 0.0 – 0.00.0 – 0.0 0.0 – 0.00.0 – 0.0 0.0 – 0.0

Qualitative Ranges N/A

Assay Configuration: Base PageReaction Mode Wavelength-Prim/Sec Read time-Main/Flex AbsMaxVarEND UP 500 / 660 31 – 33 / 0 – 0 0.0Sample Blank Test Blank Read Time Abs Window Abs Limits_____ ( ___ ) 0 – 0 0 – 0 0.0 – 0.0

S.Vol DS.Vol D.Vol W.VolStandard 2.4 0.0 0 0 Rgt Name/PosDil 1 25.0 2.4 75 0 Diluent: DILUENT D–18*Dil 2 2.4 0.0 0 0 Type# 0

Rgt Name/Pos R.Vol W.Vol Type#Reagent 1 CHOL061 – ___* 240 0 0Reaction Check Read Time – A/B Range Minimum___________ 1 – 1 / 1 – 1 0.0 – 0.0 0.0Factor/Intercept Decimal Places Units

1.0 / 0.0 0 mg/dL

Assay Configuration: Calibration PageCalib Mode Interval (H)Linear 720Blank/Calib Replicates Extrapolation % Span Span Abs Range3 / 3 0 BLK – 1 0.0 – 0.0

Sample S.Vol DS.Vol D.Vol W.Vol Blk Abs RangeBLK Water 2.4 0.0 0 0 0.0 – 0.0C1 MCC 1 2.4 0.0 0 0 Cal DeviationC2 MCC 2 2.4 0.0 0 0 0.0

FAC Limit (%)10

Assay Configuration: SmartWash PageRgt Probe

Reagent Wash VolALBP061 Water 300

CuvetteAssay Name Wash Vol— — —

Sample ProbeWash—

Assay Configuration: Outline PageAssay Name Assay # LineChol 18 B-LineQuantitative RangesMin Text Min Panic-L L-Reference-H Panic-H Max Max Text

* 0.0* 0.0 0.00 5.17 0.0 0.0* *

0.17** L-Linear Range-H 18.26

Reference Ranges*Age Male Female

0 Year0 Year0 Year

0.0 – 0.0 0.0 – 0.00.0 – 0.0 0.0 – 0.00.0 – 0.0 0.0 – 0.00.0 – 0.0 0.0 – 0.0


Assay Configuration: Base PageReaction Mode Wavelength-Prim/Sec Read time-Main/Flex AbsMaxVarEND UP 500 / 660 31 – 33 / 0 – 0 0.0Sample Blank Test Blank Read Time Abs Window Abs Limits_____ ( ___ ) 0 – 0 0 – 0 0.0 – 0.0

S.Vol DS.Vol D.Vol W.VolStandard 2.4 0.0 0 0 Rgt Name/PosDil 1 25.0 2.4 75 0 Diluent: DILUENT D–18*Dil 2 2.4 0.0 0 0 Type# 0

Rgt Name/Pos R.Vol W.Vol Type#Reagent 1 CHOL061 – ___* 240 0 0Reaction Check Read Time – A/B Range Minimum___________ 1 – 1 / 1 – 1 0.0 – 0.0 0.0Factor/Intercept Decimal Places Units

1.0 / 0.0 2 mmol/L

Assay Configuration: Calibration PageCalib Mode Interval (H)Linear 720Blank/Calib Replicates Extrapolation % Span Span Abs Range3 / 3 0 BLK – 1 0.0 – 0.0

Sample S.Vol DS.Vol D.Vol W.Vol Blk Abs RangeBLK Water 2.4 0.0 0 0 0.0 – 0.0C1 MCC 1 2.4 0.0 0 0 Cal DeviationC2 MCC 2 2.4 0.0 0 0 0.0

FAC Limit (%)10

Assay Configuration: SmartWash PageRgt Probe

Reagent Wash VolALBP061 Water 300

CuvetteAssay Name Wash Vol— — —

Sample ProbeWash—

1

ULTRA HDL

This package insert contains information to run the Ultra HDL assay on the ARCHITECT c Systems™ and the

AEROSET System.


NOTE: This package insert must be read carefully prior to product use. Package insert instructions must be

followed accordingly. Reliability of assay results cannot be guaranteed if there are any deviations from the

instructions in this package insert.

Customer Support

United States: 1-877-4ABBOTT

Canada: 1-800-387-8378 (English speaking customers)

1-800-465-2675 (French speaking customers)

International: Call your local Abbott representative

Symbols in Product Labeling

Calibrator Catalog number/List number

Concentration Serial number

Authorized Representative in the European Community


Ingredients Manufacturer

In vitro diagnostic medical device Temperature limitation

Batch code/Lot number Use by/Expiration date

Reagent 1

Reagent 2

ULTRA HDL

3K33-2030-4129/R5

ABBOTT LABORATORIESAbbott Park, IL 60064, USA


May 2008©2005, 2008 Abbott Laboratories

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NAMEULTRA HDL

INTENDED USEThe Ultra HDL (UHDL) assay is used for the quantitation of high density lipoprotein (HDL) cholesterol in human serum or plasma.

SUMMARY AND EXPLANATION OF TESTPlasma lipoproteins are spherical particles containing varying amounts of cholesterol, triglycerides, phospholipids, and proteins. Phospholipids, free cholesterol, and proteins constitute the outer surface of the lipoprotein particle, while the inner core contains mostly esterified cholesterol and triglyceride. These particles serve to solubilize and transport cholesterol and triglyceride in the bloodstream.

The relative proportions of protein and lipid determine the density of these lipoproteins and provide a basis on which to begin their classification.1 The classes are: chylomicron, very-low-density lipoprotein (VLDL), low-density lipoprotein (LDL), and high-density lipoprotein (HDL). Numerous clinical studies have shown that the different lipoprotein classes have very distinct and varied effects on coronary heart disease risk.2

The principle role of HDL cholesterol in lipid metabolism is the uptake and transport of cholesterol from peripheral tissues to the liver through a process known as reverse cholesterol transport (a proposed cardioprotective mechanism).3 Low HDL cholesterol levels are strongly associated with an increased risk of coronary heart disease.4-7 Hence, the determination of serum HDL cholesterol is a useful tool in identifying high-risk patients. The Adult Treatment Panel of the National Cholesterol Education Program (NCEP) recommends that in all adults 20 years of age and over, a fasting lipoprotein profile (total cholesterol, LDL cholesterol, HDL cholesterol, and triglyceride) should be obtained once every five years to screen for coronary heart disease risk.8

PRINCIPLES OF PROCEDUREThe Ultra HDL assay is a homogeneous method for directly measuring HDL cholesterol concentrations in serum or plasma without the need for off-line pretreatment or centrifugation steps.

The method uses a two-reagent format and depends on the properties of a unique detergent. This method is based on accelerating the reaction of cholesterol oxidase (CO) with non-HDL unesterified cholesterol and dissolving HDL cholesterol selectively using a specific detergent. In the first reagent, non-HDL unesterified cholesterol is subject to an enzyme reaction and the peroxide generated is consumed by a peroxidase reaction with DSBmT yielding a colorless product. The second reagent consists of a detergent (capable of solubilizing HDL cholesterol), cholesterol esterase (CE), and chromagenic coupler to develop color for the quantitative determination of HDL cholesterol.

Methodology: Accelerator Selective Detergent

REAGENTS

Reagent Kit

3K33 Ultra HDL is supplied as a liquid, ready-to-use, two-reagent kit which contains:

4 x 84 mL4 x 32 mL

Estimated tests per kit: 1,440 Calculation is based on the minimum reagent fill volume per kit.

Reactive Ingredients Concentration

Cholesterol oxidase (E. coli) < 1,000 U/L

Peroxidase (Horseradish) < 1,300 ppg U/L

N, N-bis (4-sulphobutyl)-m-toluidine-disodium (DSBmT)

< 1.0 mmol/L

Accelerator < 1.0 mmol/L

Ascorbic oxidase (Curcubita sp.) < 3,000 U/L

Cholesterol esterase (Pseudomonas sp.) < 1,500 U/L

4-Aminoantipyrine < 0.1%

Detergent < 2.0%

The Ultra HDL reagent is certified as traceable to the HDL cholesterol designated comparison method, covering the NCEP medical decision points, by the CDC-Certified Cholesterol Reference Method Laboratory Network (CRMLN).


Reagent Handling

Remove air bubbles, if present in the reagent cartridge, with a new applicator stick. Alternatively, allow the reagent to sit at the appropriate storage temperature to allow the bubbles to dissipate. To minimize volume depletion, do not use a transfer pipette to remove the bubbles.

CAUTION: Reagent bubbles may interfere with proper detection of reagent level in the cartridge, causing insufficient reagent aspiration which could impact results.

Reagent Storage

• Unopened reagents are stable until the expiration date when stored at 2 to 8°C.

• DO NOT FREEZE.• Protect reagents from direct sunlight.• Reagent stability is 28 days if the reagent is uncapped and onboard.

Indications of Deterioration

• Quality control results outside of the acceptance criteria defined by your laboratory.

• Presence of turbidity.


Precautions for Users

1. For in vitro diagnostic use.2. Do not use components beyond the expiration date.3. Do not mix materials from different kit lot numbers.4. CAUTION: This product requires the handling of human specimens.

It is recommended that all human sourced materials be considered potentially infectious and be handled in accordance with the OSHA Standard on Bloodborne Pathogens.8 Biosafety Level 29 or other appropriate biosafety practices10,11 should be used for materials that contain or are suspected of containing infectious agents.

5. and contain a mixture of 5-chloro-2-methyl-4-isothiazolin-3-one and 2-methyl-4-isothiazolin-3-one (3:1), which is a component of ProClin, and are classified per applicable European Community (EC) Directives as: Irritant (Xi). The following are the appropriate Risk (R) and Safety (S) phrases:

R43 May cause sensitization by skin contact.S24 Avoid contact with skin.S35 This material and its container must be disposed

of in a safe way.S37 Wear suitable gloves.S46 If swallowed, seek medical advice immediately

and show this container or label.


Suitable Specimens

Serum and plasma are acceptable specimens. The National Cholesterol Education Program (NCEP) recommends using fasting specimens for a lipoprotein profile. If the specimen is nonfasting, only the values for total cholesterol and HDL cholesterol are usable.12

• Serum: Use serum collected by standard venipuncture techniques into glass or plastic tubes with or without gel barriers. Ensure complete clot formation has taken place prior to centrifugation. When processing samples, separate serum from blood cells or gel according to the specimen collection tube manufacturer’s instructions.Some specimens, especially those from patients receiving anticoagulant or thrombolytic therapy, may take longer to complete their clotting processes. Fibrin clots may subsequently form in these sera and the clots could cause erroneous test results.

• Plasma: Use plasma collected by standard venipuncture techniques into glass or plastic tubes. Acceptable anticoagulants are sodium heparin, lithium heparin (with or without gel barrier), and spray-dried EDTA.* Ensure centrifugation is adequate to remove platelets. When processing samples, separate plasma from blood cells or gel according to the specimen collection tube manufacturer’s instructions.

*NOTE: Lower HDL cholesterol results obtained from EDTA plasma have been attributed to an osmotic dilution effect. The NCEP has suggested multiplying EDTA plasma results by a factor of 1.03 to correct the EDTA result to a serum equivalent value.13


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SPECIMEN COLLECTION AND HANDLING (Continued)

Specimen Storage

Serum and Plasma

Temperature MaximumStorage


20 to 25°C 2 days 14

2 to 8°C 7 days 14, 15

-20°C 3 months 14



PROCEDURE

Materials Provided

3K33 Ultra HDL Reagent Kit

Materials Required but not Provided

• 1E68 HDL Calibrator, 6 x 1 mL• Control Material• Saline (0.85% to 0.90% NaCl) for specimens that require dilution

Assay Procedure

For a detailed description of how to run an assay, refer to Section 5 of the instrument-specific operations manual.

Specimen Dilution Procedures

The ARCHITECT c Systems and the AEROSET System have automatic dilution features; refer to Section 2 of the instrument-specific operations manual for additional information.

Serum and plasma: Specimens with HDL cholesterol values exceeding 180 mg/dL (4.66 mmol/L) are flagged and may be diluted using the Automated Dilution Protocol or the Manual Dilution Procedure.

Automated Dilution Protocol

If using the Automated Dilution Protocol, the system performs a dilution of the specimen and automatically corrects the concentration by multiplying the result by the appropriate dilution factor. To set up the automatic dilution feature, refer to Section 2 of the instrument-specific operations manual for additional information.

Manual Dilution Procedure

Manual dilutions should be performed as follows:

• Use saline (0.85% to 0.90% NaCl) to dilute the sample.• The operator must enter the dilution factor in the patient or control

order screen. The system uses this dilution factor to automatically correct the concentration by multiplying the result by the entered factor.

• If the operator does not enter the dilution factor, the result must be multiplied by the appropriate dilution factor before reporting the result.



CALIBRATIONCalibration is stable for approximately 28 days (672 hours) and is required with each change in reagent lot number. Verify calibration with at least two levels of controls according to the established quality control requirements for your laboratory. If control results fall outside acceptable ranges, recalibration may be necessary.

For a detailed description of how to calibrate an assay, refer to Section 6 of the instrument-specific operations manual.

For information on calibrator standardization, refer to the HDL Calibrator package insert.

QUALITY CONTROLThe following is the recommendation of Abbott Laboratories for quality control. As appropriate, refer to your laboratory standard operating procedure(s) and/or quality assurance plan for additional quality control requirements and potential corrective actions.• Two levels of controls (normal and abnormal) are to be run every

24 hours.• If more frequent control monitoring is required, follow the established

quality control procedures for your laboratory.• If quality control results do not meet the acceptance criteria

defined by your laboratory, patient values may be suspect. Follow the established quality control procedures for your laboratory. Recalibration may be necessary.

• Review quality control results and acceptance criteria following a change of reagent or calibrator lot.


• ARCHITECT System Operations Manual—Appendix C• AEROSET System Operations Manual—Appendix ARepresentative performance data are given in the EXPECTED VALUES and SPECIFIC PERFORMANCE CHARACTERISTICS sections of this package insert. Results obtained in individual laboratories may vary.

LIMITATIONS OF THE PROCEDUREUsing three homogenous HDL assays, Camps, et al. have reported artificially low HDL results in patients with liver cirrhosis.16 Published studies are not available that define the severity of liver disease necessary to affect lipoprotein and HDL metabolism, or establish other possible patterns of interference with HDL results. When an HDL result is diagnostically critical with concomitant clinically relevant liver disease, use a recognized precipitation or ultracentrifugation HDL-reference method for confirmation. Artificially decreased or increased HDL values in the presence of dyslipidemias have been reported.17,18

Refer to the SPECIMEN COLLECTION AND HANDLING and SPECIFIC PERFORMANCE CHARACTERISTICS sections of this package insert.

For the AEROSET System only: Ultra HDL must be run on a separate line from 7D60-02 Total Bilirubin (TBil) and 7D74-20 Triglyceride (Trig).

EXPECTED VALUES

Reference Range

Serum/Plasma12

Range(mg/dL)

Range(mmol/L)

Major risk factor for heart disease < 40 < 1.04

Negative risk factor for heart disease ≥ 60 ≥ 1.55

To convert results from mg/dL to mmol/L, multiply mg/dL by 0.0259.The National Cholesterol Education Program (NCEP) Adult Treatment Panel III Report recommends the classification shown above. Laboratories should follow recommendations for lipid ranges effective in their locale if they differ from those of the NCEP.


Linearity

Ultra HDL is linear up to 180 mg/dL (4.66 mmol/L), with recovery within 10% of the predicted value with 95% confidence.Linearity was verified using a modified Clinical and Laboratory Standards Institute (CLSI) protocol NCCLS EP6-A.19 An internal verification study produced linear results up to 221 mg/dL (5.72 mmol/L).

Limit of Detection and Quantitation

The limit of quantitation (LOQ) for Ultra HDL is 5.0 mg/dL (0.13 mmol/L), and the limit of detection (LOD) is 2.5 mg/dL (0.06 mmol/L).

The LOD testing for Ultra HDL was performed using a study design based on CLSI protocol NCCLS EP17-A.20 An internal verification study produced an LOD for Ultra HDL of 0.3 mg/dL (0.01 mmol/L). The proportions of false positives (α) and false negatives (β) were less than 5% and the limit of blank (LOB) was 0.2 mg/dL (0.01 mmol/L).

The LOQ is the analyte concentration at which the CV = 20%. An internal verification study produced a CV of 9.1% at an HDL cholesterol concentration of 4.4 mg/dL (0.11 mmol/L).

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(Continued)

Interfering Substances

Interference studies were conducted using an acceptance criteria of 5% of the target value. Interference effects were assessed by Dose Response method, at the medical decision levels of the analyte.

Lower Decision Level

Interfering Substance Interferent Concentration N

Target(mg/dL)


Ascorbic Acid 2.9 mg/dL (165 μmol/L) 3 35 99

3.9 mg/dL (221 μmol/L) 3 35 99

Conjugated Bilirubin

32.6 mg/dL (557 μmol/L) 3 34 104

63.3 mg/dL (1,082 μmol/L) 3 34 77

Unconjugated Bilirubin

32.4 mg/dL (554 μmol/L) 3 33 105

65.5 mg/dL (1,120 μmol/L) 3 33 107

Hemoglobin 1,000 mg/dL (10 g/L) 3 31 102

2,000 mg/dL (20 g/L) 3 31 104

Intralipid 1,000 mg/dL (10 g/L) 3 32 102

2,000 mg/dL (20 g/L) 3 32 115

Upper Decision Level

Interfering Substance Interferent Concentration N

Target(mg/dL)


Ascorbic Acid 2.9 mg/dL (165 μmol/L) 3 69 101

3.9 mg/dL (221 μmol/L) 3 69 101

Conjugated Bilirubin

32.0 mg/dL (547 μmol/L) 3 68 102

63.5 mg/dL (1,086 μmol/L) 3 68 95

Unconjugated Bilirubin

33.9 mg/dL (580 μmol/L) 3 67 102

67.1 mg/dL (1,147 μmol/L) 3 67 102

Hemoglobin 1,000 mg/dL (10 g/L) 3 62 99

2,000 mg/dL (20 g/L) 3 62 100

Intralipid 1,000 mg/dL (10 g/L) 3 75 99

2,000 mg/dL (20 g/L) 3 75 101

Ascorbic acid solutions at the above concentrations were prepared by addition of L-ascorbic acid to human serum pools. Conjugated bilirubin solutions at the above concentrations were prepared by addition of a ditaurobilirubin stock to human serum pools. Unconjugated bilirubin solutions at the above concentrations were prepared by addition of a NIST SRM 916a bilirubin stock to human serum pools. Hemoglobin solutions at the above concentrations were prepared by addition of hemolysate to human serum pools. Intralipid solutions at the above concentrations were prepared by addition of Intralipid to human serum pools.

Interferences from medications or endogenous substances may affect results.21

Precision

The imprecision of the Ultra HDL assay is total SD ≤ 1.7 mg/dL or total CV ≤ 4%, whichever is greater. Internal verification studies were performed using CLSI protocol NCCLS EP5-A.22 Representative data are summarized below.

Control Level 1 Level 2

N 80 80

Mean (mg/dL) 20.9 78.9

Within RunSD 0.36 0.76

%CV 1.7 1.0

Between RunSD 0.23 0.36

%CV 1.1 0.5

Between DaySD 1.07 0.73

%CV 5.1 0.9

TotalSD 1.15 1.11

%CV 5.5 1.4

Accuracy

Accuracy data for Ultra HDL were collected using the HDL Cholesterol Certification Protocol for Manufacturers.23 The data were analyzed using CLSI protocol NCCLS EP21-A.24

Serum results from the Ultra HDL assay on an ARCHITECT c System and an AEROSET System were compared with the designated comparison method (DCM) for HDL cholesterol.

ARCHITECT AEROSET

Mean %Bias -1.6 -1.8

%Total Error 10.9 10.2

Method Comparison

Correlation studies were performed using CLSI protocol NCCLS EP9-A2.25

Serum results from the Ultra HDL assay on the AEROSET System were compared with those from a commercially available accelerator selective detergent methodology.

Serum results from the Ultra HDL assay on an ARCHITECT c System were compared with those from the Ultra HDL assay on an AEROSET System.

AEROSET vs. Comparative Method

ARCHITECT vs. AEROSET

N 111 110

Y - Intercept 0.46 0.61

Correlation Coefficient 0.999 0.999

Slope 0.97 1.00

%Bias at 35 mg/dL -2 1

%Bias at 60 mg/dL -2 1

Range (mg/dL) 12 to 188 12 to 179

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BIBLIOGRAPHY 1. Gotto AM. Lipoprotein metabolism and the etiology of

hyperlipidemia. Hosp Pract 1988;23(Suppl 1):4–13. 2. Third Report of the National Cholesterol Education Program (NCEP)

Expert Panel on Detection, Evaluation, and Treatment of High Blood Cholesterol in Adults (Adult Treatment Panel III)—Final Report. National Institutes of Health. National Heart, Lung, and Blood Institute. NIH Publication No. 02-5215. September 2002; I-1–II-22.

3. Badimon JJ, Badimon L, Fuster V. Regression of atherosclerotic lesions by high density lipoprotein plasma fraction in the cholesterol-fed rabbit. J Clin Invest 1990;85(4):1234–41.

4. Castelli WP, Doyle JT, Gordon T, et al. HDL Cholesterol and other lipids in coronary heart disease. The cooperative lipoprotein phenotyping study. Circulation 1977;55(5):767–72.

5. Gordon T, Castelli WP, Hjortland MC, et al. High density lipoprotein as a protective factor against coronary heart disease. Am J Med 1977;62(5):707.

6. Williams P, Robinson D, Bailey A. High density lipoprotein and coronary risk factors in normal men. Lancet 1979;1(8107):72–5.

7. Kannel WB, Castelli WP, Gordon T. Cholesterol in the prediction of atherosclerotic disease; New perspectives based on the Framingham study. Ann Intern Med 1979;90(1):85–91.

8. US Department of Labor, Occupational Safety and Health Administration. 29 CFR Part 1910.1030. Occupational Exposure to Bloodborne Pathogens.

9. US Department of Health and Human Services. Biosafety in Microbiological and Biomedical Laboratories, 5th ed. Washington, DC: US Government Printing Office; January 2007.

10. World Health Organization. Laboratory Biosafety Manual, 3rd ed. Geneva: World Health Organization; 2004.

11. Sewell DL, Bove KE, Callihan DR, et al. Protection of Laboratory Workers from Occupationally Acquired Infections; Approved Guideline―Third Edition (M29-A3). Wayne, PA: Clinical and Laboratory Standards Institute, 2005.

12. Executive summary of the third report of the National Cholesterol Education Program (NCEP) Expert Panel on detection, evaluation, and treatment of high blood cholesterol in adults (Adult Treatment Panel III). JAMA 2001;285(19):2486–97.

13. National Cholesterol Education Program. Recommendations on Lipoprotein Measurement, from the Working Group on Lipoprotein Measurement. National Institutes of Health. National Heart, Lung, and Blood Institute. NIH Publication No. 95-3044. September 1995.

14. Guder WG, da Fonseca-Wollheim F, Heil W, et al. The Quality of Diagnostic Samples. Darmstadt, Germany: GIT Verlag; 2001:22–3.

15. US Pharmacopeial Convention, Inc. General notices. In: US Pharmacopeia National Formulary, 1995 ed (USP 23/NF 18). Rockville, MD: The US Pharmacopeial Convention, Inc; 1994:11.

16. Camps J, Simo JM, Guaita S, et al. Altered Composition of Lipoproteins in Liver Cirrhosis Compromises Three Homogenous Methods for HDL-Cholesterol. Clin Chem 1999;45(5):685–88.

17. Roberts WL, Leary ET, Lambert TL, et al. Falsely low direct HDL-cholesterol results in a patient with dysbetalipoproteinemia. Clin Chem 2000;46:560–2.

18. Lackner, KJ, Schmitz G. Beta-VLDL of patients with type III hyperlipoproteinemia interferes with homogenous determination of HDL-cholesterol based on polyethylene glycol-modified enzymes. Clin Chem 1998;44:2546–8.

19. Tholen DW, Kroll M, Astles JR, et al. Evaluation of the Linearity of Quantitative Measurement Procedures: A Statistical Approach; Approved Guideline (EP6-A). Wayne, PA: The National Committee for Clinical Laboratory Standards, 2003.

20. Tholen DW, Linnet K, Kondratovich M, et al. Protocols for Determination of Limits of Detection and Limits of Quantitation; Approved Guideline (EP17-A). Wayne, PA: The National Committee for Clinical Laboratory Standards, 2004.

21. Young DS, Effects of Drugs on Clinical Laboratory Tests, 5th ed. Washington, DC: AACC Press, 2000:3-399–3-414.

22. Kennedy JW, Carey RN, Coolen RB, et al. Evaluation of Precision Performance of Clinical Chemistry Devices; Approved Guideline (EP5-A). Wayne, PA: The National Committee for Clinical Laboratory Standards, 1999.

23. Cholesterol Reference Method Laboratory Network. HDL Cholesterol Certification Protocol for Manufacturers. November 2002. Accessed July 11, 2005 from: http://www.cdc.gov/labstandards/pdf/crmln/MFRHDLNov2002final.pdf.

24. Krouwer JS, Astles JR, Cooper WG, et al. Estimation of Total Analytical Error for Clinical Laboratory Methods; Approved Guideline (EP21-A). Wayne, PA: The National Committee for Clinical Laboratory Standards, 2003.

25. Krouwer JS, Tholen DW, Garber CC, et al. Method Comparison and Bias Estimation Using Patient Samples; Approved Guideline—Second Edition (EP9-A2). Wayne, PA: The National Committee for Clinical Laboratory Standards, 2002.

TRADEMARKSAEROSET and ARCHITECT are registered trademarks of Abbott Laboratories.

c System is a trademark of Abbott Laboratories.

All other trademarks, brands, product names, and trade names are the property of their respective companies.

Licensed under PCT/JP00/03860 and PCT/JP97/04442.

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* User defined.† Due to differences in instrument systems and unit configurations, version numbers may vary.‡ Displays the number of decimal places defined in the decimal places parameter field.†† Refer to concentration specified on calibrator labeling.

Ultra HDL Serum/Plasma—Conventional and SI Units

Ultra HDL Serum/Plasma—Conventional Units

Ultra HDL Serum/Plasma—SI Units

ARCHITECT c SYSTEMS ASSAY PARAMETERS


● General о Calibration о SmartWash о Results о InterpretationAssay: UHDL Type: Photometric Version: †

Number: 1093

● Reaction definition о Reagent / Sample о Validity checksReaction mode: End up

Primary Secondary Read timesWavelength: 604 / 700 Main: 31 – 33

Last required read: 33

Absorbance range: ___ – ___ Color correction: ___ – ___Sample blank type: Self Blank: 14 – 16

о Reaction definition ● Reagent / Sample о Validity checksR1 R2

Reagent: UHDL0 Reagent volume: 200 67

Diluent: Saline Water volume: ___ ___Diluent dispense mode:Type 0 Dispense mode: Type 0 Type 0

Diluted DefaultDilution name Sample sample Diluent Water Dilution factor dilutionSTANDARD : 2.0 ___ ___ ___ = 1:1.00 ●_________ : ___ ___ ___ ___ = о_________ : ___ ___ ___ ___ = о


Maximum absorbance variation: ___


о General ● Calibration о SmartWash о Results о InterpretationAssay: UHDL Calibration method: Linear

● Calibrators о Volumes о Intervals о Validity checksCalibrator set: Calibrator level: Concentration:

UHDL Blank: Water 0‡

Cal 1: UHDL1 ††Replicates: 3 [Range 1 – 3]

о Calibrators ● Volumes о Intervals о Validity checksCalibrator: UHDL Diluted

Calibrator level Sample sample Diluent WaterBlank: Water 2.0 ___ ___ ___Cal 1: UHDL1 2.0 ___ ___ ___


Full interval: 672 (hours)Calibration type:

Adjust type: None

о Calibrators о Volumes о Intervals ● Validity checksBlank absorbance range: _____ – _____

Span: Blank – Blank

Span absorbance range: _____ – _____Expected cal factor: 0.00

Expected cal factor tolerance %: 0

Configure assay parameters — c 8000 SmartWash

о General о Calibration ● SmartWash о Results о Interpretation

Assay: UHDL

COMPONENT REAGENT / ASSAY WASH Volume ReplicatesR1 TRIG0 10% Detergent B*** 345 1

Cuvette Trig 10% Detergent B 345

*** Select “Detergent B” for software prior to version 2.2.

Configure assay parameters — Results

о General о Calibration о SmartWash ● Results о InterpretationAssay: UHDL Assay number: 1093

Dilution default range: Result units: mg/dL

Low-Linearity: 5

High-Linearity: 180

Gender and age specific ranges:*GENDER AGE (UNITS) NORMAL EXTREMEEither 0 – 130 (Y) 40 – 60

Configure result units

Assay: UHDL

Version: †Result units: mg/dL


Intercept: 0.0000


о General о Calibration о SmartWash ● Results о InterpretationAssay: UHDL Assay number: 1093

Dilution default range: Result units: mmol/L

Low-Linearity: 0.13

High-Linearity: 4.66

Gender and age specific ranges:*GENDER AGE (UNITS) NORMAL EXTREMEEither 0 – 130 (Y) 1.04 – 1.55


Assay: UHDL

Version: †Result units: mmol/L


Intercept: 0.0000

Configure assay parameters — c 16000 SmartWash


Assay: UHDL

COMPONENT REAGENT / ASSAY WASH Volume ReplicatesR1 AlbG0 Detergent A 345 1

R1 AlbP0 Water 345 1

R1 TRIG0 10% Detergent B 345 1


7

Refer to Assay Configuration in Section 2 of the AEROSET System Operations Manual for information regarding assay parameters.* User defined or instrument defined.** Ultra HDL must be run on a separate line from 7D60-02 Total Bilirubin (TBil) and 7D74-20 Triglyceride (Trig).

Ultra HDL Serum/Plasma—Conventional Units

AEROSET SYSTEM ASSAY PARAMETERS

Assay Configuration: Outline Page

Assay Name Assay # Line

UHDL 93 A-Line**Quantitative Ranges

Min Text Min Panic-L L-Reference-H Panic-H Max Max Text

* 0.0* 0.0 40 60 0.0 0.0* *

5 L-Linear Range-H 180

Reference Ranges*

Age Male Female

0 Year0 Year0 Year

0.0 – 0.0 0.0 – 0.00.0 – 0.0 0.0 – 0.00.0 – 0.0 0.0 – 0.00.0 – 0.0 0.0 – 0.0


Assay Configuration: Base Page

Reaction Mode Wavelength-Prim/Sec Read time-Main/Flex AbsMaxVar

END UP 604 / 700 31 – 33 / 0 – 0 0.0Sample Blank Test Blank Read Time Abs Window Abs Limits

UHDL ( 93 ) 14 – 16 0 – 0 0.0 – 0.0S.Vol DS.Vol D.Vol W.Vol

Standard 2.0 0.0 0 0 Rgt Name/Pos

Dil 1 2.0 0.0 0 0 Diluent: _______ – __*

Dil 2 2.0 0.0 0 0 Type# 0Rgt Name/Pos R.Vol W.Vol Type#

Reagent 1 UHDL061 – ___* 200 0 0Reagent 2 UHDL052 – ___* 67 0 0Reaction Check Read Time – A/B Range Minimum

___________ 1 – 1 / 1 – 1 0.0 – 0.0 0.0Factor/Intercept Decimal Places Units

1.0 / 0.0 0 mg/dL

Assay Configuration: Calibration Page

Calib Mode Interval (H)

Linear 672Blank/Calib Replicates Extrapolation % Span Span Abs Range

3 / 3 0 BLK – 1 0.0 – 0.0Sample S.Vol DS.Vol D.Vol W.Vol Blk Abs Range

BLK Water 2.0 0.0 0 0 0.0 – 0.0C1 UHDL 1 2.0 0.0 0 0 Cal Deviation

0.0FAC Limit (%)

10

Assay Configuration: SmartWash Page

Rgt Probe

Reagent Wash Vol

ALBG061 AlkW 345ALBP061 AlkW 345

Cuvette

Assay Name Wash Vol

— — —Sample Probe

Wash

—

Ultra HDL Serum/Plasma—SI Units



UHDL 93 A-Line**Quantitative Ranges


* 0.0* 0.0 1.04 1.55 0.0 0.0* *

0.13 L-Linear Range-H 4.66

Reference Ranges*

Age Male Female

0 Year0 Year0 Year

0.0 – 0.0 0.0 – 0.00.0 – 0.0 0.0 – 0.00.0 – 0.0 0.0 – 0.00.0 – 0.0 0.0 – 0.0



Reaction Mode Wavelength-Prim/Sec Read time-Main/Flex AbsMaxVar

END UP 604 / 700 31 – 33 / 0 – 0 0.0Sample Blank Test Blank Read Time Abs Window Abs Limits

UHDL ( 93 ) 14 – 16 0 – 0 0.0 – 0.0S.Vol DS.Vol D.Vol W.Vol


Dil 1 2.0 0.0 0 0 Diluent: _______ – __*

Dil 2 2.0 0.0 0 0 Type# 0Rgt Name/Pos R.Vol W.Vol Type#

Reagent 1 UHDL061 – ___* 200 0 0Reagent 2 UHDL052 – ___* 67 0 0Reaction Check Read Time – A/B Range Minimum

___________ 1 – 1 / 1 – 1 0.0 – 0.0 0.0Factor/Intercept Decimal Places Units

1.0 / 0.0 2 mmol/L



Linear 672Blank/Calib Replicates Extrapolation % Span Span Abs Range

3 / 3 0 BLK – 1 0.0 – 0.0Sample S.Vol DS.Vol D.Vol W.Vol Blk Abs Range

BLK Water 2.0 0.0 0 0 0.0 – 0.0C1 UHDL 1 2.0 0.0 0 0 Cal Deviation

0.0FAC Limit (%)

10


Rgt Probe

Reagent Wash Vol

ALBG061 AlkW 345ALBP061 AlkW 345

Cuvette

Assay Name Wash Vol

— — —Sample Probe

Wash

—

1/9

CRP VarioNOTE: This package insert must be read carefully prior to product use. Package insert instructions must be followed accordingly. Reliability of assay results cannot be guaranteed if there are any deviations from the instructions in this package insert.


INTENDED USEThe MULTIGENT CRP Vario assay [CRPVa] is intended for the quantitative immunoturbidimetric determination of C-reactive protein in human serum and plasma with variable assay ranges [CRP16, CRP32, CRP48] using the ARCHITECT c Systems and the AEROSET System.

SUMMARY AND EXPLANATION OF TESTC-reactive protein (CRP) is an acute phase protein whose concentration rises non-specifically in response to inflammation. CRP is seen to increase as a result of the inflammatory process, most notably in response to pneumococcal (bacterial) infection, histolytic disease, and a variety of other disease states. Intraindividual variation is a major limitation of the assay when the assay is used for directing therapies. Intraindividual variations of the CRP levels are from 30% to 60%. Serial measurement may be required to estimate true mean of CRP depending on the intended use in any specific individual. CRP is used as a marker or general diagnostic indicator of infections and inflammation, in addition to serving as a monitor of patient response to pharmacological therapy and surgery.

PRINCIPLES OF PROCEDURECRP Vario is a latex immunoassay developed to accurately and reproducibly measure blood CRP levels in serum and plasma. When an antigen-antibody reaction occurs between CRP in a sample and anti-CRP antibody, which has been adsorbed to latex particles, agglutination results. This agglutination is detected as an absorbance change (572 nm), with the rate of change being proportional to the quantity of CRP in the sample. Three different methods (High Sensitivity [CRP16], Standard [CRP32], and Wide Range [CRP48]) are available to cover a wide analytical measurement range.

Methodology: Turbidimetric/Immunoturbidimetric

REAGENTS

Reagent Kit

6K26-40 MULTIGENT CRP Vario is supplied as a two-reagent kit, which contains:

3 x 90 mL

3 x 90 mL

Method Estimated Tests per Kit*

High Sensitivity 2,192Standard 2,192Wide Range 1,843

*Calculation is based on the minimum reagent fill volume per kit.

Reactive Ingredients Concentration

Glycine buffer (pH 7.0) 1.28%Rabbit anti-CRP polyclonal antibodies adsorbed on latex particles

0.2%

Inactive Ingredients: contains bovine albumin (≤ 1%) and sodium azide (< 0.1%). contains bovine albumin (≤ 0.1%) and sodium azide (< 0.1%).


Reagent Handling

• Ready for use.• Ready for use.• Remove air bubbles, if present in the reagent cartridge, with a

new applicator stick. Alternatively, allow the reagent to sit at the appropriate storage temperature to allow the bubbles to dissipate. To minimize volume depletion, do not use a transfer pipette to remove the bubbles.

CAUTION: Reagent bubbles may interfere with proper detection of reagent level in the cartridge, causing insufficient reagent aspiration that could impact results.

Reagent Storage

• Unopened reagents are stable until the expiration date when stored at 2 to 8°C.

• Reagent stability is 60 days if the reagent is uncapped and onboard.

Indications of Deterioration

Instability or deterioration should be suspected if there are visible signs of leakage, extreme turbidity, microbial growth, if calibration does not meet the appropriate package insert and/or instrument-specific operations manual criteria, or if controls do not meet the appropriate criteria.


Precautions for Users

• For in vitro diagnostic use.• Do not use components beyond the expiration date.• Do not mix materials from different kit lot numbers.• CAUTION: This product requires the handling of human specimens.

It is recommended that all human sourced materials be considered potentially infectious and be handled in accordance with the OSHA Standard on Bloodborne Pathogens.1 Biosafety Level 22 or other appropriate biosafety practices3,4 should be used for materials that contain or are suspected of containing infectious agents.

• This product contains sodium azide; for a specific listing, refer to the REAGENTS section of this package insert. Contact with acids liberates very toxic gas. This material and its container must be disposed of in a safe way.NOTE: Refer to Section 8 of the instrument-specific operations manual for proper handling and disposal of reagents containing sodium azide.

For product not classified as dangerous per European Directive 1999/45/EC as amended – Safety data sheet available for professional user on request.


Suitable Specimens

• Serum: Use serum collected by standard venipuncture techniques into plastic tubes with or without gel. Ensure complete clot formation has taken place prior to centrifugation. When processing samples, separate serum from blood cells or gel according to the specimen collection tube manufacturer’s instructions.

Some specimens, especially those from patients receiving anticoagulant or thrombolytic therapy, may take longer to complete their clotting processes. Fibrin clots may subsequently form in these sera and the clots could cause erroneous test results.

• Plasma: Use plasma collected by standard venipuncture techniques into plastic tubes. Acceptable anticoagulants are lithium heparin (with or without gel barrier), sodium heparin, and EDTA. Ensure centrifugation is adequate to remove platelets. When processing samples, separate plasma from blood cells or gel according to the specimen collection tube manufacturer’s instructions.

NOTE: Glass tubes were not tested.


Specimen Storage

TemperatureMaximumStorage


20 to 25°C 15 days 52 to 8°C 2 months 5, 6-20°C 3 years 5



PROCEDURE

Materials Provided

6K26-40 MULTIGENT CRP Vario Kit

Materials Required but not Provided• 6K26-13 MULTIGENT CRP Calibrator Set 7 x 2 mL• 6K26-14 MULTIGENT CRP Calibrator HS 1 x 2 mL• 6K26-15 MULTIGENT CRP Calibrator WR 1 x 2 mL• 6K26-21 MULTIGENT CRP Control HS 3 x 2 mL• Saline (0.85% to 0.90% NaCl) for specimens that require dilution

Assay Procedure

For a detailed description of how to run an assay on an ARCHITECT c System or the AEROSET System, refer to Section 5 of the instrument-specific operations manual.

Specimen Dilution Procedure

The ARCHITECT c Systems and the AEROSET System have automatic dilution features; refer to Section 2 of the instrument-specific operations manual for additional information.

Serum and Plasma: Specimens with CRP values exceeding the linearity are flagged and may be diluted by following either the Automated Dilution Protocol or the Manual Dilution Procedure.

6K26-40

6K2640-3.0/ 02 2010/04/28 en

6K2640-3.0/ 02 2/9

PROCEDURE (Continued)

Specimen Dilution Procedure (Continued)

Automated Dilution Protocol

If using the Automated Dilution Protocol, the system performs a dilution of the specimen and automatically corrects the concentration by multiplying the result by the appropriate dilution factor. The dilution for each method is listed below.

Method Dilution

High Sensitivity 1:10Standard 1:5Wide Range 1:5

Manual Dilution Procedure

• Use saline (0.85% to 0.90% NaCl) to dilute the sample.

• The operator must enter the dilution factor in the patient or control order screen. The system uses this dilution factor to automatically correct the concentration by multiplying the result by the entered factor.

• If the operator does not enter the dilution factor, the result must be multiplied by the appropriate dilution factor before reporting the result.



CALIBRATIONNOTE: The MULTIGENT CRP Vario assay must be calibrated using the individual levels listed in the instrument-specific assay parameters. Refer to the parameters for the High Sensitivity [CRP16], Standard [CRP32], and Wide Range [CPR48] methods and the CRP Calibrator package insert specific for the method used in your laboratory.

Calibration is stable for approximately 15 days (360 hours) and is required with each change in reagent lot number. Verify calibration curve with at least two levels of controls according to the established quality control requirements for your laboratory. If control results fall outside acceptable ranges, recalibration may be necessary.

For a detailed description of how to calibrate an assay, refer to Section 6 of the instrument-specific operations manual.

Standardization

For information on calibrator standardization, refer to the CRP Calibrator package insert specific for the method used in your laboratory.

QUALITY CONTROLAs appropriate, refer to your laboratory standard operating procedure(s) and/or quality assurance plan for additional quality control requirements and potential corrective actions:

• Two levels of controls (normal and abnormal) are to be run every 24 hours.

• If more frequent control monitoring is required, follow the established quality control procedures for your laboratory.

• If quality control results do not meet the acceptance criteria defined by your laboratory, patient values may be suspect. Follow the established quality control procedures for your laboratory. Recalibration may be necessary.

• Review quality control results and acceptance criteria following a change of reagent or calibrator lot.


• ARCHITECT System Operations Manual—Appendix C

• AEROSET System Operations Manual—Appendix A

To convert results from mg/dL to mg/L, multiply mg/dL by 10.7

Representative performance data are given in the EXPECTED VALUES and SPECIFIC PERFORMANCE CHARACTERISTICS sections of this package insert. Results obtained in individual laboratories may vary.

LIMITATIONS OF THE PROCEDUREThe following are limitations on the use of the High Sensitivity CRP perCDC/AHA recommendations.8

• Screening the entire adult population is not recommended.

• CRP is not a substitute for traditional cardiovascular risk factors.

• Acute coronary syndrome management should not depend on CRP measurements.

• Patients with persistently unexplained CRP levels above 1.0 mg/dL (10 mg/L) should be evaluated for noncardiovascular etiologies.

• Secondary prevention measures should not depend on CRP.

• Serial measurements of CRP should not be used to monitor treatment.

• The average of two CRP results, repeated optimally two weeks apart, should be used on metabolically stable patients.

• In very rare cases gammopathy, particularly of the monoclonal IgM type (e.g., Waldenström macroglobulinemia), may cause unreliable results.

Refer to the SPECIMEN COLLECTION AND HANDLING and SPECIFIC PERFORMANCE CHARACTERISTICS sections of this package insert.

EXPECTED VALUES

Reference RangeRange (mg/dL) Range (mg/L)

Serum and plasma9 ≤ 0.5 ≤ 5

Schlebusch et al.10 have published pediatric reference ranges.

CRP is an acute phase protein whose concentration rises non-specifically in response to inflammation. CRP values should not be interpreted without a complete clinical evaluation. Follow-up testing of patients with elevated values is recommended in order to help rule out a recent response to undetected infection or tissue injury. It is recommended that each laboratory establish its own expected range. For diagnostic purposes, the patient’s medical history and all other clinical findings should be considered when evaluating CRP results.


Reportable Range

The reportable range for MULTIGENT CRP Vario is:

High Sensitivity Method 0.01 to 16.00 mg/dL (0.1 to 160 mg/L}Standard Method 0.02 to 32.00 mg/dL (0.2 to 320 mg/L)Wide Range 0.02 to 48.00 mg/dL (0.2 to 480 mg/L)

All three methods were tested for prozone up to a CRP concentration of 100 mg/dL (1,000 mg/L). No prozone effect was observed within the linear range of the assay. At 100 mg/dL (1,000 mg/L) the observed result was correctly flagged as above the linearity of the assay.

Limit of Quantitation (LOQ)

The LOQ is the analyte concentration at which the CV = 20%. The limit of quantification for MULTIGENT CRP Vario is:

High Sensitivity Method 0.01 mg/dL (0.1 mg/L)Standard and Wide Range Methods 0.02 mg/dL (0.2 mg/L)

Interfering Substances

Interference studies were conducted using an acceptance criteria of ± 5% deviation from the target value.

Interfering Substance Interferent Concentration

Bilirubin, conjugated and fetal 30 mg/dL (513 μmol/L)Hemoglobin 500 mg/dL (5 g/L)Intralipid 1,500 mg/dL (15 g/L)Rheumatoid factor 550 IU/mL (550 kU/L)

Precision

Precision was determined over five days with two runs and four replicates of each control per day. Representative results in mg/L are summarized below.

CRP High Sensitivity Method

Control Level 1 Level 2 Level 3 Level 4

N 40 40 40 40Mean (mg/L) 0.6 5.0 14.9 53.0

Within RunSD 0.022 0.07 0.13 0.26

%CV 3.62 1.32 0.86 0.50

Between RunSD 0.005 0.02 0.10 0.33

%CV 0.85 0.38 0.64 0.62

TotalSD 0.022 0.07 0.16 0.40

%CV 3.72 1.46 1.04 0.76

CRP Standard Method

Control Level 1 Level 2 Level 3

N 40 40 40Mean (mg/L) 5.0 14.9 53.8

Within RunSD 0.06 0.14 0.50

%CV 1.11 0.93 0.92

Between RunSD 0.04 0.13 0.47

%CV 0.80 0.89 0.87

TotalSD 0.07 0.18 0.64

%CV 1.41 1.21 1.19

CRP Wide Range Method

Control Level 1 Level 2 Level 3

N 40 40 40Mean (mg/L) 5.4 18.3 263.8

Within RunSD 0.07 0.17 2.99

%CV 1.29 0.93 1.13

Between RunSD 0.05 0.13 2.65

%CV 0.92 0.71 1.00

TotalSD 0.08 0.22 4.02

%CV 1.48 1.18 1.52

3/9

SPECIFIC PERFORMANCE CHARACTERISTICS (Continued)

Method Comparison

Serum results from the MULTIGENT CRP Vario methods on the AEROSET System were compared with the results from a commercially available nephelometric methodology.

Serum results from the MULTIGENT CRP Vario methods on an ARCHITECT c System were compared with the results on the AEROSET System.

For the MULTIGENT CRP High Sensitivity method only, serum results from an ARCHITECT c System and the AEROSET System were also compared with the results from a commercially available turbidimetric methodology.

Method comparison data are presented in mg/L.

CRP High Sensitivity MethodAEROSET vs.

Nephelometer

ARCHITECT vs.

AEROSET

N 45 45Y - Intercept (95% CI*) -0.24 to 0.54 -0.33 to 0.45Correlation Coefficient 0.9994 0.9994Slope (95% CI*) 0.985 to 1.006 0.975 to 0.996Standard Error of the Estimate 0.96 0.96

Range (mg/L) 1.0 to 104.0 1.1 to 103.3

* CI = Confidence Interval

CRP High Sensitivity Method

(continued)

AEROSET vs.

Turbidimetric

Method

ARCHITECT vs.

Turbidimetric

Method


Range (mg/L) 0.2 to 13.6 0.2 to 13.6


CRP Standard MethodAEROSET vs.

Nephelometer

ARCHITECT vs.

AEROSET


Range (mg/L) 1.0 to 219.0 0.6 to 223


CRP Wide Range MethodAEROSET vs.

Nephelometer

ARCHITECT vs.

AEROSET


Range (mg/L) 1.0 to 286.0 0.7 to 301.2


BIBLIOGRAPHY1. US Department of Labor, Occupational Safety and Health Administration.

29 CFR Part 1910.1030. Bloodborne Pathogens.

2. US Department of Health and Human Services. Biosafety in Microbiological and Biomedical Laboratories, 5th ed. Washington, DC: US Government Printing Office, January 2007.

3. World Health Organization. Laboratory Biosafety Manual, 3rd ed. Geneva: World Health Organization, 2004.

4. Sewell DL, Bove KE, Callihan DR, et al. Protection of Laboratory Workers from Occupationally Acquired Infections; Approved Guideline—Third Edition (M29-A3). Wayne, PA: Clinical and Laboratory Standards Institute, 2005.

5. Guder WG, da Fonseca-Wollheim F, Heil W, et al. The Quality of Diagnostic Samples. Darmstadt, Germany: GIT Verlag; 2001:24–5.

6. US Pharmacopeial Convention, Inc. General notices. In: US Pharmacopeia National Formulary, 1995 ed (USP 23/NF 18). Rockville, MD: The US Pharmacopeial Convention, Inc; 1994:11.

7. Burtis CA, Ashwood ER, Bruns DE, editors. Tietz Textbook of Clinical Chemistry and Molecular Diagnostics, 4th ed. St Louis, MO: Elsevier Saunders; 2006:2263.

8. Pearson TA, Mensah GA, Alexander RW, et al. Markers of inflammation and cardiovascular disease: application to clinical and public health practice: A statement for healthcare professionals from the Centers for Disease Control and Prevention and the American Heart Association. Circulation 2003;107(3):499–511.

9. Dati F, Johnson AM, Whicher JT. The existing interim consensus reference ranges and the future approach. Clin Chem Lab Med 2001;39(11):1134–6.

10. Schlebusch H, Liappis N, Kalina E, et al. High sensitive CRP and creatinine: reference intervals from infancy to childhood. J Lab Med 2002;26(5/6):341–6.

TRADEMARKSCRP Vario is a trademark of Sentinel CH. SpA in various jurisdictions.

The ARCHITECT c System family of instruments consists of c 4000, c 8000, and c 16000 Systems.

AEROSET, ARCHITECT, c 4000, c 8000, c 16000, c System, and MULTIGENT are trademarks of Abbott Laboratories in various jurisdictions.

All other trademarks are property of their respective owners.

US Patent: 6,248,597 / 6,828,158 and equivalent patents in other countries.

SYMBOLS IN PRODUCT LABELING

Calibrator

Contents of kit

Control

In vitro diagnostic medical device

Batch code/Lot number

Reagent 1

Reagent 2

Catalog number/List number

Serial number


Use by/Expiration date

Manufacturer

Temperature limitation

CONTACT INFORMATIONFor product questions contact Abbott Laboratories Customer Support.

United States:Canada:

International:

1-877-4ABBOTT1-800-387-8378 (English speaking customers)1-800-465-2675 (French speaking customers)Call your local Abbott representative

SENTINEL CH. SpAVia Robert Koch, 2Milan 20152 Italy

Distributed by:Abbott Laboratories Inc.

Abbott Park, IL 60064 USA

6K2640-3.0/ 02 4/9

† Due to differences in instrument systems and unit configurations, version numbers may vary.‡ Refer to the concentration specified on calibrator labeling. In ARCHITECT software version 5.00 and above, these values are defined on the

Configure calibrator set screen.‡‡ Short name does not display on instrument screen.* User defined.

c Systems Assay Parameters

CRP Vario (High Sensitivity Method) Serum/Plasma:

Conventional and SI Units


Conventional Units


● General о Calibration о SmartWash о Results о InterpretationAssay: CRP16 Type: Photometric Version: †

Number: 2974

● Reaction definition о Reagent / Sample о Validity checksReaction mode: Rate up

Primary Secondary Read times

Wavelength: 572 / None Main: 20 – 26

Last required read: 26 Flex: ___ – ___

Absorbance range: 0.7000 – 3.2000 Color correction: ___ – ___

Sample blank type: None


Reagent: CRP0S Reagent volume: 100 100

Diluent: Saline Water volume: ___ ___

Diluent dispense mode: Type 0 Dispense mode: Type 0 Type 0

Diluted DefaultDilution name Sample sample Diluent Water Dilution factor dilution

STANDARD : 4.0 ___ ___ ___ = 1:1.00 ●DIL 1 : 10.0 4.0 90 ___ = 1:10.00 о________ : ___ ___ ___ ___ = ___ о


Rate linearity %: ___


Assay: CRP16

Version: †

Result units: mg/dL

Decimal places: 2 [Range 0 – 4]

Correlation factor: 1.0000

Intercept: 0.0000


SI Units


Assay: CRP16

Version: †

Result units: mg/L



Intercept: 0.0000



Assay: CRP16

COMPONENT REAGENT / ASSAY WASH Volume Replicates



о General ● Calibration о SmartWash о Results о InterpretationAssay: CRP16 Calibration method: Spline

● Calibrators о Volumes о Intervals о Validity checks

Calibrator set: Calibrator level: (Short name)‡‡Concentration:‡

(mg/dL) (mg/L)

16CRP Blank: Water 0.00 0.00

Cal 1: 16CRP1 (CRPHS) 0.25 2.50

Replicates: 2 [Range 1 - 3] Cal 2: 16CRP2 (CRP05) 0.50 5.00

Cal 3: 16CRP3 (CRP10) 1.00 10.00

Cal 4: 16CRP4 (CRP20) 2.00 20.00

Cal 5: 16CRP5 (CRP80) 8.00 80.00

Cal 6: 16CRP6 (CRP160) 16.00 160.00

о Calibrators ● Volumes о Intervals о Validity checksCalibrator: 16CRP Diluted

Calibrator level Sample sample Diluent Water

Blank: Water 4.0 ___ ___ ___

Cal 1: 16CRP1 4.0 ___ ___ ___

Cal 2: 16CRP2 4.0 ___ ___ ___

Cal 3: 16CRP3 4.0 ___ ___ ___

Cal 4: 16CRP4 4.0 ___ ___ ___

Cal 5: 16CRP5 4.0 ___ ___ ___

Cal 6: 16CRP6 4.0 ___ ___ ___


Full interval: 360 (hours)

Calibration type:

Adjust type: None

о Calibrators о Volumes о Intervals ● Validity checksBlank absorbance range: -0.0500 – 0.0500


Span absorbance range: _____ – _____

Expected cal factor: 0.00



о General о Calibration о SmartWash ● Results о InterpretationAssay: CRP16 Assay number: 2974


Low-Linearity: 0.01


Gender and age specific ranges:*

GENDER AGE (UNITS) NORMAL EXTREME

Either 0 – 130 (Y) 0.00 – 0.50



Dilution default range: Result units: mg/L

Low-Linearity: 0.10




Either 0 – 130 (Y) 0.00 – 5.00

5/9 6K2640-3.0/ 02


CRP Vario (Standard Method) Serum/Plasma:



Conventional Units



Number: 2973












STANDARD : 2.0 ___ ___ ___ = 1:1.00 ●DIL 1 : 20.0 2.0 80 ___ = 1:5.00 о________ : ___ ___ ___ ___ = ___ о




Assay: CRP32

Version: †

Result units: mg/dL



Intercept: 0.0000


SI Units


Assay: CRP32

Version: †

Result units: mg/L



Intercept: 0.0000



Assay: CRP32






Low-Linearity: 0.02




Either 0 – 130 (Y) 0.00 – 0.50




Low-Linearity: 0.2




Either 0 – 130 (Y) 0.0 – 5.0





(mg/dL) (mg/L)


Cal 1: 32CRP1 (CRP05) 0.50 5.0


Cal 3: 32CRP3 (CRP20) 2.00 20.0

Cal 4: 32CRP4 (CRP40) 4.00 40.0

Cal 5: 32CRP5 (CRP160) 16.00 160.0

Cal 6: 32CRP6 (CRP320) 32.00 320.0



Blank: Water 2.0 ___ ___ ___

Cal 1: 32CRP1 2.0 ___ ___ ___

Cal 2: 32CRP2 2.0 ___ ___ ___

Cal 3: 32CRP3 2.0 ___ ___ ___

Cal 4: 32CRP4 2.0 ___ ___ ___

Cal 5: 32CRP5 2.0 ___ ___ ___

Cal 6: 32CRP6 2.0 ___ ___ ___



Calibration type:

Adjust type: None








6K2640-3.0/ 02 6/9


CRP Vario (Wide Range Method) Serum/Plasma:



Conventional Units



Number: 2975












STANDARD : 2.0 ___ ___ ___ = 1:1.00 ●DIL 1 : 20.0 2.0 80 ___ = 1:5.00 о________ : ___ ___ ___ ___ = ___ о




Assay: CRP48

Version: †

Result units: mg/dL



Intercept: 0.0000


SI Units


Assay: CRP48

Version: †

Result units: mg/L



Intercept: 0.0000



Assay: CRP48






Low-Linearity: 0.02




Either 0 – 130 (Y) 0.00 – 0.50




Low-Linearity: 0.2




Either 0 – 130 (Y) 0.0 – 5.0





(mg/dL) (mg/L)


Cal 1: 48CRP1 (CRP05) 0.50 5.0


Cal 3: 48CRP3 (CRP20) 2.00 20.0

Cal 4: 48CRP4 (CRP40) 4.00 40.0

Cal 5: 48CRP5 (CRP160) 16.00 160.0

Cal 6: 48CRP6 (CRPWR) 48.00 480.0



Blank: Water 2.0 ___ ___ ___

Cal 1: 48CRP1 2.0 ___ ___ ___

Cal 2: 48CRP2 4.0 ___ ___ ___

Cal 3: 48CRP3 5.0 ___ ___ ___

Cal 4: 48CRP4 5.0 ___ ___ ___

Cal 5: 48CRP5 3.0 ___ ___ ___

Cal 6: 48CRP6 2.0 ___ ___ ___



Calibration type:

Adjust type: None








7/9 6K2640-3.0/ 02

Refer to Assay Configuration in Section 2 of the AEROSET System Operations Manual for information regarding assay parameters.* User or instrument defined.


Conventional Units


SI Units



CRP16 974 *

Quantitative Ranges


* 0.0* 0.0 0.00 5.00 0.0 0.0* *


Reference Ranges*

Age Male Female

0 Year

0 Year

0 Year

0.0 – 0.0 0.0 – 0.0

0.0 – 0.0 0.0 – 0.0

0.0 – 0.0 0.0 – 0.0

0.0 – 0.0 0.0 – 0.0



Rgt Probe

Reagent Wash Vol

— — —

Cuvette

Assay Name Wash Vol

— — —

Sample Probe

Wash

—


Reaction Mode Wavelength-Prim/Sec Read time-Main/Flex Linearity%

RATE UP 572 / – 20 – 26 / 0 – 0 0

Sample Blank Test Blank Read Time Abs Window Abs Limits

_____ ( ___ ) 0 – 0 0 – 0 0.7 – 3.2

S.Vol DS.Vol D.Vol W.Vol


Dil 1 10.0 4.0 90 0 Diluent: _____ _–__*

Dil 2 4.0 0.0 0 0 Type# 0

Rgt Name/Pos R.Vol W.Vol Type#

Reagent 1 CRP0S61 – ___* 100 0 0

Reagent 2 CRP0S62 – ___* 100 0 0

Reaction Check Read Time – A/B Range Minimum

___________ 1 – 1 / 1 – 1 0.0 – 0.0 0.0

Factor/Intercept Decimal Places Units

1.0 / 0.0 2 mg/L



CRP16 974 *

Quantitative Ranges


* 0.0* 0.0 0.00 0.50 0.0 0.0* *


Reference Ranges*

Age Male Female

0 Year

0 Year

0 Year

0.0 – 0.0 0.0 – 0.0

0.0 – 0.0 0.0 – 0.0

0.0 – 0.0 0.0 – 0.0

0.0 – 0.0 0.0 – 0.0




RATE UP 572 / – 20 – 26 / 0 – 0 0


_____ ( ___ ) 0 – 0 0 – 0 0.7 – 3.2



Dil 1 10.0 4.0 90 0 Diluent: _____ _–__*

Dil 2 4.0 0.0 0 0 Type# 0


Reagent 1 CRP0S61 – ___* 100 0 0

Reagent 2 CRP0S62 – ___* 100 0 0


___________ 1 – 1 / 1 – 1 0.0 – 0.0 0.0


1.0 / 0.0 2 mg/dL


Rgt Probe

Reagent Wash Vol

— — —

Cuvette

Assay Name Wash Vol

— — —

Sample Probe

Wash

—



Spline 360

Blank/Calib Replicates Extrapolation % Span Span Abs Range

2 / 2 0 BLK – 1 0.0 – 0.0

Sample Conc S.Vol DS.Vol D.Vol W.Vol Blk Abs Range

BLK Water 0 4.0 0.0 0 0 -0.05 – 0.05

C1 CRPHS 0.25 4.0 0.0 0 0 Cal Deviation

C2 CRP05 0.5 4.0 0.0 0 0 0.01

C3 CRP10 1.0 4.0 0.0 0 0 FAC Limit (%)

C4 CRP20 2.0 4.0 0.0 0 0 10

C5 CRP40 4.0 4.0 0.0 0 0

C6 CRP80 8.0 4.0 0.0 0 0

C7 CRP160 16.0 4.0 0.0 0 0



Spline 360


2 / 2 0 BLK – 1 0.0 – 0.0


BLK Water 0 4.0 0.0 0 0 -0.05 – 0.05

C1 CRPHS 2.5 4.0 0.0 0 0 Cal Deviation

C2 CRP05 5 4.0 0.0 0 0 0.01

C3 CRP10 10 4.0 0.0 0 0 FAC Limit (%)

C4 CRP20 20 4.0 0.0 0 0 10

C5 CRP40 40 4.0 0.0 0 0

C6 CRP80 80 4.0 0.0 0 0

C7 CRP160 160 4.0 0.0 0 0

6K2640-3.0/ 02 8/9



Conventional Units


SI Units



CRP32 973 *

Quantitative Ranges


* 0.0* 0.0 0.0 5.0 0.0 0.0* *


Reference Ranges*

Age Male Female

0 Year

0 Year

0 Year

0.0 – 0.0 0.0 – 0.0

0.0 – 0.0 0.0 – 0.0

0.0 – 0.0 0.0 – 0.0

0.0 – 0.0 0.0 – 0.0



Rgt Probe

Reagent Wash Vol

— — —

Cuvette

Assay Name Wash Vol

— — —

Sample Probe

Wash

—



RATE UP 572 / – 20 – 26 / 0 – 0 0


_____ ( ___ ) 0 – 0 0 – 0 0.7 – 3.2



Dil 1 20.0 2.0 80 0 Diluent: _____ _–__*

Dil 2 2.0 0.0 0 0 Type# 0


Reagent 1 CRP0S61 – ___* 100 0 0

Reagent 2 CRP0S62 – ___* 100 0 0


___________ 1 – 1 / 1 – 1 0.0 – 0.0 0.0


1.0 / 0.0 1 mg/L



CRP32 973 *

Quantitative Ranges


* 0.0* 0.0 0.00 0.50 0.0 0.0* *


Reference Ranges*

Age Male Female

0 Year

0 Year

0 Year

0.0 – 0.0 0.0 – 0.0

0.0 – 0.0 0.0 – 0.0

0.0 – 0.0 0.0 – 0.0

0.0 – 0.0 0.0 – 0.0




RATE UP 572 / – 20 – 26 / 0 – 0 0


_____ ( ___ ) 0 – 0 0 – 0 0.7 – 3.2



Dil 1 20.0 2.0 80 0 Diluent: _____ _–__*

Dil 2 2.0 0.0 0 0 Type# 0


Reagent 1 CRP0S61 – ___* 100 0 0

Reagent 2 CRP0S62 – ___* 100 0 0


___________ 1 – 1 / 1 – 1 0.0 – 0.0 0.0


1.0 / 0.0 2 mg/dL


Rgt Probe

Reagent Wash Vol

— — —

Cuvette

Assay Name Wash Vol

— — —

Sample Probe

Wash

—



Spline 360


2 / 2 0 BLK – 1 0.0 – 0.0


BLK Water 0 2.0 0.0 0 0 -0.05 – 0.05

C1 CRP05 0.5 2.0 0.0 0 0 Cal Deviation

C2 CRP10 1.0 2.0 0.0 0 0 0.01

C3 CRP20 2.0 2.0 0.0 0 0 FAC Limit (%)

C4 CRP40 4.0 2.0 0.0 0 0 10

C5 CRP80 8.0 2.0 0.0 0 0

C6 CRP160 16.0 2.0 0.0 0 0

C7 CRP320 32.0 2.0 0.0 0 0



Spline 360


2 / 2 0 BLK – 1 0.0 – 0.0


BLK Water 0 2.0 0.0 0 0 -0.05 – 0.05

C1 CRP05 5 2.0 0.0 0 0 Cal Deviation

C2 CRP10 10 2.0 0.0 0 0 0.01

C3 CRP20 20 2.0 0.0 0 0 FAC Limit (%)

C4 CRP40 40 2.0 0.0 0 0 10

C5 CRP80 80 2.0 0.0 0 0

C6 CRP160 160 2.0 0.0 0 0

C7 CRP320 320 2.0 0.0 0 0

9/9 6K2640-3.0/ 02


** The calibrator concentrations in this column must be used for this method to correctly calculate the calibration curve.NOTE: Do not use the concentrations listed on the calibrator vial labels for this method.

CRP Vario (Wide Range Method) Serum/Plasma—

Conventional Units

CRP Vario (Wide Range Method) Serum/Plasma—SI Units



CRP48 975 *

Quantitative Ranges


* 0.0* 0.0 0.0 5.0 0.0 0.0* *


Reference Ranges*

Age Male Female

0 Year

0 Year

0 Year

0.0 – 0.0 0.0 – 0.0

0.0 – 0.0 0.0 – 0.0

0.0 – 0.0 0.0 – 0.0

0.0 – 0.0 0.0 – 0.0



Rgt Probe

Reagent Wash Vol

— — —

Cuvette

Assay Name Wash Vol

— — —

Sample Probe

Wash

—



RATE UP 572 / – 20 – 26 / 0 – 0 0


_____ ( ___ ) 0 – 0 0 – 0 0.7 – 3.2



Dil 1 20.0 2.0 80 0 Diluent: _____ _–__*

Dil 2 2.0 0.0 0 0 Type# 0


Reagent 1 CRP0S61 – ___* 120 0 0

Reagent 2 CRP0S62 – ___* 120 0 0


___________ 1 – 1 / 1 – 1 0.0 – 0.0 0.0


1.0 / 0.0 1 mg/L



CRP48 975 *

Quantitative Ranges


* 0.0* 0.0 0.00 0.50 0.0 0.0* *


Reference Ranges*

Age Male Female

0 Year

0 Year

0 Year

0.0 – 0.0 0.0 – 0.0

0.0 – 0.0 0.0 – 0.0

0.0 – 0.0 0.0 – 0.0

0.0 – 0.0 0.0 – 0.0




RATE UP 572 / – 20 – 26 / 0 – 0 0


_____ ( ___ ) 0 – 0 0 – 0 0.7 – 3.2



Dil 1 20.0 2.0 80 0 Diluent: _____ _–__*

Dil 2 2.0 0.0 0 0 Type# 0


Reagent 1 CRP0S61 – ___* 120 0 0

Reagent 2 CRP0S62 – ___* 120 0 0


___________ 1 – 1 / 1 – 1 0.0 – 0.0 0.0


1.0 / 0.0 2 mg/dL


Rgt Probe

Reagent Wash Vol

— — —

Cuvette

Assay Name Wash Vol

— — —

Sample Probe

Wash

—



Spline 360


2 / 2 0 BLK – 1 0.0 – 0.0

Sample Conc** S.Vol DS.Vol D.Vol W.Vol Blk Abs Range

BLK Water 0 2.0 0.0 0 0 -0.05 – 0.05

C1 CRP05 5 2.0 0.0 0 0 Cal Deviation

C2 CRP10 20 4.0 0.0 0 0 0.01

C3 CRP20 50 5.0 0.0 0 0 FAC Limit (%)

C4 CRP40 100 5.0 0.0 0 0 10

C5 CRP160 240 3.0 0.0 0 0

C6 CRPWR 480 2.0 0.0 0 0



Spline 360


2 / 2 0 BLK – 1 0.0 – 0.0

Sample Conc** S.Vol DS.Vol D.Vol W.Vol Blk Abs Range

BLK Water 0 2.0 0.0 0 0 -0.05 – 0.05

C1 CRP05 0.5 2.0 0.0 0 0 Cal Deviation

C2 CRP10 2.0 4.0 0.0 0 0 0.01

C3 CRP20 5.0 5.0 0.0 0 0 FAC Limit (%)

C4 CRP40 10.0 5.0 0.0 0 0 10

C5 CRP160 24.0 3.0 0.0 0 0

C6 CRPWR 48.0 2.0 0.0 0 0

1

system

enTSH

7K62

49-3481/R3B7K620

Read Highlighted ChangesRevised June, 2010

TSH

Customer Service: Contact your local representative or find country specific contact information on www.abbottdiagnostics.com.

Package insert instructions must be carefully followed. Reliability of assay results cannot be guaranteed if there are any deviations from the instructions in this package insert.

Key to symbols used

List Number

In Vitro Diagnostic Medical Device

Serial Number

Lot Number

Expiration Date

Store at 2-8°C


Manufacturer

Reagent Lot

Calibrator (1, 2)

Control Low, Medium, High (L, M, H)

Assay CD-ROM

Reaction Vessels

Sample Cups

Septum

Replacement Caps

Warning: Severe Irritant

See REAGENTS section for a full explanation of symbols used in reagent component naming.

2

NAMEARCHITECT TSH

INTENDED USEThe ARCHITECT TSH assay is a Chemiluminescent Microparticle Immunoassay (CMIA) for the quantitative determination of human thyroid stimulating hormone (TSH) in human serum and plasma.

SUMMARY AND EXPLANATION OF TEST Human Thyroid Stimulating Hormone (TSH) or thyrotropin is a glycoprotein with a molecular weight of approximately 28,000 daltons, synthesized by the basophilic cells (thyrotropes) of the anterior pituitary.1 TSH is composed of two non-covalently linked subunits designated alpha and beta. Although the alpha subunit of TSH is common to the luteinizing hormone (LH), follicle stimulating hormone (FSH) and human chorionic gonadotropin (hCG), the beta subunits of these glycoproteins are hormone specific and confer biological as well as immunological specificity. Both alpha and beta subunits are required for biological activity.1 TSH stimulates the production and secretion of the metabolically active thyroid hormones, thyroxine (T4) and triiodothyronine (T3), by interacting with a specific receptor on the thyroid cell surface.2 T3 and T4 are responsible for regulating diverse biochemical processes throughout the body which are essential for normal development and metabolic and neural activity.The synthesis and secretion of TSH is stimulated by thyrotropin releasing hormone (TRH), the hypothalamic tripeptide, in response to low levels of circulating thyroid hormones.3,4 Elevated levels of T3 and T4 suppress the production of TSH via a classic negative feedback mechanism. Other evidence also indicates that somatostatin and dopamine exert inhibitory control over TSH release, suggesting that the hypothalamus may provide both inhibitory and stimulatory influence on pituitary TSH production.5

Failure at any level of regulation of the hypothalamic-pituitary-thyroid axis will result in either underproduction (hypothyroidism) or overproduction (hyperthyroidism) of T4 and/or T3.In cases of primary hypothyroidism, T3 and T4 levels are low and TSH levels are significantly elevated.6 In the case of pituitary dysfunction, either due to intrinsic hypothalamic or pituitary disease; i.e., central hypothyroidism, normal or marginally elevated basal TSH levels are often seen despite significant reduction in T4 and/or T3 levels. These inappropriate TSH values are due to a reduction in TSH bioactivity which is frequently observed in such cases. Routine TRH stimulation is advised to confirm the diagnosis in such cases. Secondary hypothyroidism typically results in an impaired TSH response to TRH, while in tertiary hypothyroidism the TSH response to TRH may be normal, prolonged or exaggerated.7-9 Primary hyperthyroidism (e.g., Grave’s Disease, nodular goiter) is associated with high levels of thyroid hormones and depressed or undetectable levels of TSH.10 The TRH stimulation test has been used in diagnosis of hyperthyroidism. Hyperthyroid patients show a subnormal response to the TRH test.11 In addition, large doses of glucocorticoids, somatostatin, dopamine and replacement doses of thyroid hormones reduce or totally blunt the TSH response to TRH.11,12

Earlier assays for serum TSH lacked the sensitivity to be used as a primary test of thyroid function.13 Sensitive TSH assays now available, with increased ability to clearly distinguish between euthyroid and hyperthyroid populations, are changing thyroid function testing. Analytical sensitivity, as a means of assessing low concentration accuracy, is being replaced by functional sensitivity.14 The American Thyroid Association has formally recommended the use of functional sensitivity as the means to quantify the sensitivity of TSH assays,15 although analytical sensitivity is still widely used. Third generation TSH assays exhibit 20% interassay CVs at < 0.02 μIU/mL and are useful in the discrimination of patients with true hyperthyroidism from those with TSH suppression seen in subclinical hyperthyroidism and some non-thyroidal illnesses.16 Other thyroid tests (Free T4 estimate, Total T4, T-Uptake, and Total T3) combined with the ability to accurately measure low levels of TSH, improve the efficiency of thyroid diagnosis.17

The ARCHITECT TSH assay is used as an aid in the assessment of thyroid status, diagnosis of thyroid disease, and treatment of thyroid disease.

BIOLOGICAL PRINCIPLES OF THE PROCEDUREThe ARCHITECT TSH assay is a two-step immunoassay to determine the presence of thyroid stimulating hormone (TSH) in human serum and plasma using Chemiluminescent Microparticle Immunoassay (CMIA) technology with flexible assay protocols, referred to as Chemiflex.

In the first step, sample, anti-β TSH antibody coated paramagnetic microparticles and TSH Assay Diluent are combined. TSH present in the sample binds to the anti-TSH antibody coated microparticles. After washing, anti-α TSH acridinium labeled conjugate is added in the second step. Pre-Trigger and Trigger Solutions are then added to the reaction mixture; the resulting chemiluminescent reaction is measured as relative light units (RLUs). A direct relationship exists between the amount of TSH in the sample and the RLUs detected by the ARCHITECT i optical system.For additional information on system and assay technology, refer to the ARCHITECT System Operations Manual, Section 3.

REAGENTSReagent Kit, 100 Tests/500 TestsNOTE: Some kit sizes are not available in all countries or for use on all ARCHITECT i Systems. Please contact your local distributor.ARCHITECT TSH Reagent Kit (7K62)• 1 or 4 Bottle(s) (6.6 mL/27.0 mL) Anti-β TSH

(mouse, monoclonal) coated Microparticles in TRIS buffer with protein (bovine) stabilizers. Preservative: antimicrobial agents.

• 1 or 4 Bottle(s) (5.9 mL/26.3 mL) Anti-α TSH (mouse, monoclonal) acridinium-labeled Conjugate in MES buffer with protein (bovine) stabilizers. Minimum concentration: 60 ng/mL. Preservative: antimicrobial agent.

• 1 or 4 Bottle(s) (8.0 mL/40.7 mL) TSH Assay Diluent in TRIS buffer. Preservative: antimicrobial agents.

Manual DiluentARCHITECT i Multi-Assay Manual Diluent (7D82-50)• 1 Bottle (100 mL) ARCHITECT

i Multi-Assay Manual Diluent containing phosphate buffered saline solution. Preservative: antimicrobial agent.

Other ReagentsARCHITECT i Pre-Trigger Solution• Pre-Trigger Solution containing 1.32%

(w/v) hydrogen peroxide. ARCHITECT i Trigger Solution• Trigger Solution containing 0.35N sodium

hydroxide. ARCHITECT i Wash BufferNOTE: Bottle and volume varies based on order.• Wash Buffer containing phosphate buffered saline

solution. Preservatives: antimicrobial agents.

WARNINGS AND PRECAUTIONS•

For • In Vitro Diagnostic UsePackage insert instructions must be carefully followed. Reliability of •assay results cannot be guaranteed if there are any deviations from the instructions in this package insert.

Safety PrecautionsCAUTION:• This product may require the handling of human specimens. It is recommended that all human sourced materials be considered potentially infectious and handled in accordance with the OSHA Standard on Bloodborne Pathogens18. Biosafety Level 219 or other appropriate biosafety practices20,21 should be used for materials that contain or are suspected of containing infectious agents.

The following warnings and precautions apply to this component:Assay Diluent•

WARNING: Contains Tris Hydroxymethyl Aminomethane and Tromethamine Hydrochloride.

H315 Causes skin irritation.H319 Causes serious eye irritation.H335 May cause respiratory irritation.PreventionP264 Wash hands thoroughly after handlingP280 Wear protective gloves / protective

clothing / eye protection.P261 Avoid breathing mist / vapours / spray.

P271 Use only outdoors or in a well-ventilated area.

3

ResponseP305+P351 + P338

IF IN EYES: Rinse cautiously with water for several minutes. Remove contact lenses, if present and easy to do. Continue rinsing.

P337+P313 If eye irritation persists: Get medical advice / attention.

P302+P352 IF ON SKIN: Wash with plenty of soap and water.

P332+P313 If skin irritation occurs: Get medical advice / attention.

P362 Take off contaminated clothing and wash before reuse.

P304+340 IF INHALED: Remove victim to fresh air and keep at rest in a position comfortable for breathing.

P312 Call a POISON CENTER or doctor/physician if you feel unwell.

StorageP403 + P233 Store in a well-ventilated place. Keep

container tightly closed.P405 Store locked up.

This material and its container must be disposed of in a safe way.For a detailed discussion of safety precautions during system •operation, refer to the ARCHITECT System Operations Manual, Section 8.

Handling PrecautionsDo not use reagent kits beyond the expiration date.•Do not mix reagents from different reagent kits.•Prior to loading the ARCHITECT TSH Reagent Kit on the system for •the first time, the microparticle bottle requires mixing to resuspend microparticles that have settled during shipment. For microparticle mixing instructions, refer to the PROCEDURE, Assay Procedure section of this package insert.Septums MUST be used to prevent reagent evaporation and •contamination and to ensure reagent integrity. Reliability of assay results cannot be guaranteed if septums are not used according to the instructions in this package insert. To avoid contamination, wear clean gloves when placing a septum •on an uncapped reagent bottle.Once a septum has been placed on an open reagent bottle, • do not invert the bottle as this will result in reagent leakage and may compromise assay results.Over time, residual liquids may dry on the septum surface. These are •typically dried salts which have no effect on assay efficacy.For a detailed discussion of handling precautions during system •operation, refer to the ARCHITECT System Operations Manual, Section 7.

Storage Instructions

• The ARCHITECT TSH Reagent Kit must be stored at 2-8°C and may be used immediately after removal from 2-8°C storage.When stored and handled as directed, reagents are stable until the •expiration date.The ARCHITECT TSH Reagent Kit may be stored on-board the •ARCHITECT i System for a maximum of 30 days. After 30 days, the reagent kit must be discarded. For information on tracking on-board time, refer to the ARCHITECT System Operations Manual, Section 5.Reagents may be stored on or off the ARCHITECT• i System. If reagents are removed from the system, store them at 2-8°C (with septums and replacement caps) in an upright position. For reagents stored off the system, it is recommended that they be stored in their original trays and boxes to ensure they remain upright. If the microparticle bottle does not remain upright (with a septum installed) while in refrigerated storage off the system, the reagent kit must be discarded. After reagents are removed from the system, you must initiate a scan to update the on-board stability timer.

Indications of Reagent DeteriorationWhen a control value is out of the specified range, it may indicate deterioration of the reagents or errors in technique. Associated test results may be invalid and may require retesting. Assay recalibration may be necessary. For troubleshooting information, refer to the ARCHITECT System Operations Manual, Section 10.

INSTRUMENT PROCEDUREThe ARCHITECT TSH assay file must be installed on the ARCHITECT• i System from the ARCHITECT i Assay CD-ROM prior to performing the assay. For detailed instructions on assay file installation, refer to the ARCHITECT System Operations Manual, Section 2.For a detailed description of system procedures, refer to the •ARCHITECT System Operations Manual.The default result unit for the ARCHITECT TSH assay is μIU/ mL. An •alternate result unit, mIU/L, may be selected for reporting results by editing assay parameter “Result concentration units”, to mIU/L. The conversion factor used by the system is 1.

SPECIMEN COLLECTION AND PREPARATION FOR ANALYSISHuman serum (including serum collected in serum separator tubes) •or plasma collected in lithium heparin, sodium heparin, or potassium EDTA anticoagulant tubes may be used in the ARCHITECT TSH assay. Other anticoagulants have not been validated for use with the ARCHITECT TSH assay.Follow these package insert instructions as well as the specimen •collection tube manufacturer’s instructions for specimen collection and preparation for analysis. Refer to the specimen collection tube manufacturer’s instructions for centrifugation time and speed.Insufficient processing of sample, or disruption of the sample •during transportation may cause depressed results.For optimal results, serum and plasma specimens should be free •of fibrin, red blood cells, or other particulate matter. Centrifuge specimens containing fibrin, red blood cells, or particulate matter prior to use to ensure consistency in the results.Ensure that • complete clot formation in serum specimens has taken place prior to centrifugation. Some specimens, especially those from patients receiving anticoagulant or thrombolytic therapy may exhibit increased clotting time. If specimens are centrifuged before a complete clot forms, the presence of fibrin or particulate matter may cause erroneous results. Centrifuge specimens containing fibrin, red blood cells, or particulate matter. Note that interfering levels of fibrin may be present in samples that do not have obvious or visible particulate matter.If proper specimen collection and preparation cannot be verified, •or if samples have been disrupted due to transportation or sample handling, an additional centrifugation step is recommended. Centrifugation conditions should be sufficient to remove particulate matter. Aliquots poured versus pipetted from specimen tube types that do not include serum separators are at higher risk of including particulates and generating depressed results. Failure to follow these instructions may result in depressed •specimen results.If testing will be delayed more than 24 hours, remove serum or •plasma from the clot, serum separator or red blood cells. Specimens may be stored for up to 7 days at 2-8°C prior to being tested. If testing will be delayed more than 7 days, specimens should be frozen at -10°C or colder. Specimens stored frozen at -10°C or colder for 6 months showed no performance difference.The ARCHITECT• i System does not provide the capability to verify specimen type. It is the responsibility of the operator to verify the correct specimen types are used in the ARCHITECT TSH assay.Use caution when handling patient specimens to prevent cross •contamination. Use of disposable pipettes or pipette tips is recommended. For optimal results, inspect all samples for bubbles. Remove bubbles •with an applicator stick prior to analysis. Use a new applicator stick for each sample to prevent cross contamination.

4

Multiple freeze-thaw cycles of specimens should be avoided. •Specimens must be mixed THOROUGHLY after thawing, by LOW speed vortexing or by gently inverting, and centrifuged prior to use to remove red blood cells or particulate matter to ensure consistency in the results.When shipped, specimens must be packaged and labeled in •compliance with applicable state, federal and international regulations covering the transport of clinical specimens and infectious substances. Specimens may be shipped ambient or under thermally controlled refrigerated conditions. Prior to shipment, it is recommended that specimens be removed from the clot, serum separator or red blood cells.

PROCEDUREMaterials Provided

7K62 ARCHITECT TSH Reagent Kit•

Materials Required but not ProvidedARCHITECT• i SystemARCHITECT• i 7K62-01 ARCHITECT TSH Calibrators•7K62-10 ARCHITECT TSH Controls•7D82-50 ARCHITECT• i Multi-Assay Manual DiluentARCHITECT• i ARCHITECT• i ARCHITECT • i ARCHITECT • i ARCHITECT • i ARCHITECT • i ARCHITECT • i For information on materials required for maintenance procedures, •refer to the ARCHITECT System Operations Manual, Section 9. Pipettes or pipette tips (optional) to deliver the volumes specified on •the patient or control order screen.

Assay ProcedureBefore loading the ARCHITECT TSH Reagent Kit on the system for •the first time, the microparticle bottle requires mixing to resuspend microparticles that have settled during shipment:

Invert the microparticle bottle 30 times.•Visually inspect the bottle to ensure microparticles are •resuspended. If microparticles are still adhered to the bottle, continue to invert the bottle until the microparticles have been completely resuspended.Once the microparticles have been resuspended, remove and •discard the cap. Wearing clean gloves, remove a septum from the bag. Carefully snap the septum onto the top of the bottle.If the microparticles do not resuspend, • DO NOT USE. Contact your local Abbott representative.

Order tests.•Load the ARCHITECT TSH Reagent Kit on the ARCHITECT• i System. Verify that all necessary assay reagents are present. Ensure that septums are present on all reagent bottles.The minimum sample cup volume is calculated by the system and •is printed on the Orderlist report. No more than 9 replicates may be sampled from the same sample cup. To minimize the effects of evaporation verify adequate sample cup volume is present prior to running the test.

Priority: 200 μL for the first TSH test plus 150 μL for each •additional TSH test from the same sample cup≤ 3 hours onboard: 200 μL for the first TSH test plus 150 μL for •each additional TSH test from the same sample cup> 3 hours onboard: additional sample volume is required. Refer •to the ARCHITECT System Operations Manual, Section 5 for information on sample evaporation and volumes.

If using primary or aliquot tubes, use the sample gauge to ensure •sufficient patient specimen is present.ARCHITECT TSH Calibrators and Controls should be mixed by gentle •inversion prior to use.To obtain the recommended volume requirements for the ARCHITECT •TSH Calibrators and Controls, hold the bottles vertically and dispense 6 drops of each calibrator or 4 drops of each control into each respective sample cup.

Load samples•For information on loading samples, refer to the ARCHITECT •System Operations Manual, Section 5

Press RUN. The ARCHITECT• i System performs the following function:

Moves the sample to the aspiration point•Loads a reaction vessel (RV) into the process path•Aspirates and transfers sample into the RV•Advances the RV one position and transfers microparticles and •diluent into the RVMixes, incubates and washes the reaction mixture•Adds conjugate to the RV•Mixes, incubates and washes the reaction mixture•Adds Pre-Trigger and Trigger Solutions•Measures chemiluminescent emission to determine the quantity •of TSH in the sampleAspirates contents of RV to liquid waste and unloads RV to solid •wasteCalculates the result•

For information on ordering patient specimens, calibrators and •controls, and general operating procedures refer to the ARCHITECT System Operations Manual, Section 5.For optimal performance, it is important to follow the routine •maintenance procedures defined in the ARCHITECT System Operations Manual, Section 9. If your laboratory requires more frequent maintenance, follow those procedures.

Specimen Dilution ProceduresSpecimens with a TSH value exceeding 100.0000 μIU/mL, are flagged with the code “>100.0000” and may be diluted with either the Automated Dilution Protocol or the Manual Dilution Procedure.

If using the Automated Dilution Protocol, the system performs •a 1:5 dilution of the specimen and automatically calculates the concentration of the diluted specimen and reports the result.Manual dilutions should be performed as follows:•

The suggested dilution for TSH is 1:10. It is recommended •dilutions not exceed 1:10.For example, to perform a 1:10 dilution, add 30 μL of the patient •specimen to 270 μL of ARCHITECT i Multi-Assay Manual Diluent (7D82-50).The operator must enter the dilution factor in the patient or •control order screen. The system will use this dilution factor to automatically calculate the concentration of the sample before dilution. This will be the reported result. The result (before dilution factor is applied) should be greater than 0.0100 μIU/mL.If the operator does not enter the dilution factor, the reported •result will be that of the diluted sample. This result (before dilution factor is applied) should be greater than 0.0100 μIU/mL.

For detailed information on ordering dilutions, refer to the ARCHITECT •System Operations Manual, Section 5.

CalibrationTo perform an ARCHITECT TSH calibration, test Calibrators 1 and •2 in duplicate. A single sample of all levels of TSH controls must be tested to evaluate the assay calibration. Ensure that assay control values are within the concentration ranges specified in the package insert. Calibrators should be priority loaded.Calibrator Range: 0.0000 - 100.0000 μIU/mL.•Once an ARCHITECT TSH calibration is accepted and stored, •all subsequent samples may be tested without further calibration unless:

A reagent kit with a new lot number is used•Controls are out of range•

For detailed information on how to perform an assay calibration, refer •to the ARCHITECT System Operations Manual, Section 6.

QUALITY CONTROL PROCEDURESThe recommended control requirement for the ARCHITECT TSH assay is a single sample of all control levels tested once every 24 hours each day of use. If the quality control procedures in your laboratory require more frequent use of controls to verify test results, follow your laboratory-specific procedures. Ensure that assay control values are within the concentration ranges specified in the package insert.

5

Verification of Assay ClaimsFor protocols to verify package insert claims, refer to the ARCHITECT System Operations Manual, Appendix B. The ARCHITECT TSH assay belongs to method group 1. The lower limit of the dynamic range is defined as the functional sensitivity of the assay.

RESULTSThe ARCHITECT TSH assay utilizes a 4 Parameter Logistic Curve fit data reduction method (4PLC, Y weighted) to generate a calibration curve.

Alternate Result UnitsThe default result unit for the ARCHITECT TSH assay is μIU/mL. •When the alternate result unit, mIU/L, is selected, the conversion factor used by the system is 1.Conversion Formula: (Concentration in μIU/mL) x (1) = mIU/L.•

FlagsSome results may contain information in the Flags field. For a •description of the flags that may appear in this field, refer to the ARCHITECT System Operations Manual, Section 5.

LIMITATIONS OF THE PROCEDURESpecimens run on the ARCHITECT TSH assay MUST be processed •according to the specimen test tube manufacturer’s instruction. Insufficient processing including deviations from recommended clotting times, centrifugation times, centrifugation speed and sample preparation techniques may cause inaccurate results.For diagnostic purposes, results should be used in conjunction with •other data; e.g., symptoms, results of other thyroid tests, clinical impressions, etc.If the TSH results are inconsistent with clinical evidence, additional •testing is suggested to confirm the result.Suspected hyperthyroidism based on low or undetectable TSH levels •should be confirmed with additional thyroid function testing along with other clinical information.Specimens from patients who have received preparations of mouse •monoclonal antibodies for diagnosis or therapy may contain human anti-mouse antibodies (HAMA). Such specimens may show either falsely elevated or depressed values when tested with assay kits which employ mouse monoclonal antibodies.22,23 Additional information may be required for diagnosis.Heterophilic antibodies in human serum can react with reagent •immunoglobulins, interfering with in vitro immunoassays.24 Patients routinely exposed to animals or animal serum products can be prone to this interference and anomalous values may be observed. Additional information may be required for diagnosis.

EXPECTED VALUESA normal range of 0.35 μIU/mL to 4.94 μIU/mL (99% confidence interval) was obtained by testing serum specimens from 549 individuals defined as normal by the AxSYM Ultrasensitive hTSH II and AxSYM Free T4 assays. It is recommended that each laboratory establish its own normal range which may be unique to the population it serves depending upon geographical, patient, dietary, or environmental factors.

SPECIFIC PERFORMANCE CHARACTERISTICSPrecisionThe ARCHITECT TSH assay is designed to have a precision of ≤ 10% (total CV). A study based on guidance from Clinical and Laboratory Standards Institute (CLSI, formerly NCCLS) document EP5-A25 was performed for the ARCHITECT TSH assay. Three buffer based panel members (1, 2 and 3) and three processed human serum based panel members (4, 5 and 6) were assayed, using two lots of reagents, in replicates of two at two separate times per day for 20 testing days. Data from this study are summarized in the following table.*

Panel Member

ReagentLot

Instru- ment n

Mean Conc. (µIU/mL)

Within Run TotalSD %CV SD %CV

1 1 1 80 0.0907 0.00160 1.8 0.00210 2.31 1 2 80 0.0879 0.00121 1.4 0.00171 1.91 2 1 80 0.0876 0.00135 1.5 0.00225 2.61 2 2 80 0.0888 0.00440 5.0 0.00469 5.32 1 1 80 5.7062 0.08187 1.4 0.12184 2.12 1 2 80 5.4750 0.09116 1.7 0.12761 2.32 2 1 80 5.5153 0.08122 1.5 0.11008 2.02 2 2 80 5.5320 0.08176 1.5 0.12501 2.33 1 1 80 28.4388 0.44471 1.6 0.82863 2.93 1 2 80 27.0156 0.76916 2.8 1.03741 3.83 2 1 80 27.2486 0.58176 2.1 0.75194 2.83 2 2 80 28.0434 0.55278 2.0 0.92480 3.34 1 1 80 0.5217 0.00655 1.3 0.00894 1.74 1 2 80 0.5024 0.00751 1.5 0.01128 2.24 2 1 80 0.4998 0.00653 1.3 0.00973 1.94 2 2 80 0.5070 0.00562 1.1 0.01156 2.35 1 1 80 2.0057 0.02380 1.2 0.03367 1.75 1 2 80 1.9318 0.02679 1.4 0.03842 2.05 2 1 80 1.9060 0.03844 2.0 0.04405 2.35 2 2 80 1.9369 0.02747 1.4 0.03499 1.86 1 1 80 16.5485 0.28856 1.7 0.38175 2.36 1 2 80 15.8935 0.27310 1.7 0.41347 2.66 2 1 80 15.9947 0.25055 1.6 0.38375 2.46 2 2 80 16.3632 0.23302 1.4 0.41631 2.5

Representative data; results in individual laboratories may vary from * these data.

RecoveryThe ARCHITECT TSH assay is designed to have a mean recovery of 100 +/- 10% when analyzing samples spiked with known amounts of TSH. TSH (spanning the dynamic range) was added to 10 aliquots of human serum. The concentration of TSH was determined using the ARCHITECT TSH assay and the resulting percent recovery was calculated.* The percent recovery of the ARCHITECT TSH assay ranged from 91.8% to 104.3% with an average of 99.4%.

Representative data; results in individual laboratories may vary from * these data.

SensitivityFunctionalFunctional sensitivity is defined as the concentration of TSH that can be measured with an interassay CV of 20%.6 The ARCHITECT TSH assay is designed to have a functional sensitivity of ≤ 0.01 μlU/mL, which meets the requirements of a third generation TSH assay.In a representative study, the functional sensitivity was calculated to be ≤ 0.0038 μIU/mL (upper 95% confidence limit of 0.0042 μIU/mL). In addition, a total %CV was calculated from the pooled data generated using two lots of reagents and two instruments. The data exhibited a functional sensitivity of ≤ 0.0036 μIU/mL (upper 95% confidence limit of 0.0038 μIU/ mL). This was determined by testing human serum and processed human serum samples ranging from 0.0007 μIU/mL to 0.2365 μIU/mL. Each sample was tested over 35 to 42 days on each of two ARCHITECT i Systems using two reagent lots with at least 10 replicates per lot per instrument. The total and interassay %CVs were calculated and plotted against the mean concentration. A reciprocal curve was fitted through the data and the functional sensitivity was estimated as the concentration corresponding to the 20% CV on the fitted curve.

6

AnalyticalThe ARCHITECT TSH assay is designed to have an analytical sensitivity of ≤ 0.0025 μIU/mL. Analytical sensitivity is defined as the concentration calculated as the mean plus two standard deviations of replicates of the ARCHITECT TSH MasterCheck Level 0 (0.0 μIU/mL). The analytical sensitivity (low-linearity) is defined in the ARCHITECT TSH assay parameters as 0.0025 μIU/mL.

Analytical SpecificityThe ARCHITECT TSH assay is designed to have an analytical specificity of < 10% cross reactivity with the following substances, at the concentration levels listed, in human serum samples containing TSH in the normal range.

FSH - ≤ 500 mIU/mL•LH - ≤ 500 mIU/mL•hCG - ≤ 200,000 mIU/mL•

InterferenceThe ARCHITECT TSH assay is designed to have a potential interference from hemoglobin, bilirubin, triglycerides and protein of ≤ 10% at the levels indicated below.

Hemoglobin - ≤ 500 mg/dL•Bilirubin - ≤ 20 mg/dL•Triglycerides - ≤ 3000 mg/dL•Protein - ≤ 2 g/dL and 12 g/dL•

Accuracy by CorrelationThe ARCHITECT TSH assay is designed to have a slope of 1.0 +/- 0.2 and a correlation coefficient (r) of ≥ 0.95 when compared to the AxSYM Ultrasensitive hTSH II assay. A study was performed where specimens were tested using the ARCHITECT TSH assay and AxSYM Ultrasensitive hTSH II assay. Data from this study were analyzed using least squares and Passing-Bablok26 regression methods and are summarized in the following table.*

Abbott ARCHITECT TSH vs. Abbott AxSYM Ultrasensitive hTSH II

MethodNumber ofSpecimens Intercept Slope

CorrelationCoefficient

Least SquaresLinear Regression 534 -0.7135 0.96 0.987Passing-Bablok Linear Regression** 534 0.0098 0.91 0.987

In this evaluation, serum specimens tested ranged from 0.0109 μIU/mL to 127.9816 μIU/mL with the ARCHITECT TSH assay.

Representative data; variables such as differences in sampling size and * sample population may impact the correlation of the assay; therefore, results in individual laboratories may vary from these data.

** A linear regression method with no special assumptions regarding the distribution of the samples and the measurement errors.26

BIBLIOGRAPHYPierce JG. The Subunits of Pituitary Thyrotropin. Their Relationship to 1. other Glycoprotein Hormones. Endocrinology 1971; 89:1331-44.Rees Smith B, Pyle GA, Petersen VB, Hall R. Interaction of 2. Thyrotropin with the Human Thyrotropin Receptor. J Endocrinol 1977; 75:391-400.Sterling K, Lazarus JH. The Thyroid and Its Control. 3. Annu Rev Physiol 1977; 39:349-71.Patel YC, Alford FP, Burger HG. The 24-Hour Plasma Thyrotropin 4. Profile. Clin Sci 1972; 43:71-7.Morley JE. Neuroendocrine Control of Thyrotropin Secretion. 5. Endocr Rev 1981; 2:396-436.Burger HG, Patel YC. The Value of Serum Thyrotropin Measurement 6. in the Diagnosis and Management of Hypothyroidism. Med J Aust 1972; 2:293-7.Petersen VB, McGregor AM, Belchetz PE, Elkeles RS, Hall R. 7. The Secretion of Thyrotropin with Impaired Biological Activity in Patients with Hypothalimic-Pituitary Disease. Clin Endocrinol 1978; 8:397-402.Faglia G, Bitensky L, Pinchera A, Ferrari C, Paracchi A, Beck-Pecooz P, 8. et al. Thyrotropin Secretion in Patients with Central Hypothyroidism: Evidence for Reduced Biological Activity of Immunoreactive Thyrotropin. J Clin Endocrinol Metab 1979; 48:989-98.Beck-Peccoz P, Amr S, Menezes-Ferreira MM, Faglia G, Weintraub BD. 9. Decreased Receptor Binding of Biologically Inactive Thyrotropin in Central Hypothyroidism. N Engl J Med 1985; 312:1085-90.Wehmann RE, Rubenstein HA, Pugeat MM, Nisula BC, Extended 10. Clinical Utility of a Sensitive and Reliable Radioimmunoassay of Thyroid-Stimulating Hormone. South Med J 1983; 76:969-76.Lauridsen UB, Deckert T, Friis TH, Kirkegaard C, Hansen JM, 11. Siersbaek-Nielsen K. Estimation of Serum Thyrotropin (TSH) and Stimulation with Thyrotropin-Releasing Hormone (TRH) in Thyroid Diseases. Acta Med Scand 1974; 196:171-6.Jackson IMD. Thyrotropin-Releasing Hormone. 12. N Engl J Med 1982; 306:145-55.Spencer CA. Clinical Uses and Limitations of Rapid TSH Assays. 13. Medical Laboratory Products 1988; 17-9.Bayer MF. Performance Criteria for Appropriate Characterization of 14. “(Highly) Sensitive” Thyrotropin Assays. Clin Chem 1987; 33:630-1.Hay ID, Bayer MF, Kaplan MM, Klee GG, Larsen PR, and Spencer CA. 15. American Thyroid Association Assessment of Current Free Thyroid Hormone and Thyrotropin Measurements and Guidelines for Future Clinical Assays. Clin Chem 1991; 37:2002-8.The National Academy of Clinical Biochemistry: Standards of 16. Laboratory Practice. Laboratory Support for the Diagnosis & Monitoring of Thyroid Disease. NACB, 1996.Hay ID, Klee GG. Linking Medical Needs and Performance Goals: 17. Clinical and Laboratory Perspectives on Thyroid Disease. Clin Chem 1993; 39:1519-24.

18. US Department of Labor, Occupational Safety and Health Administration, 29 CFR Part 1910.1030, Bloodborne Pathogens.

19. US Department of Health and Human Services. Biosafety in Microbiological and Biomedical Laboratories. 5th ed. Washington, DC: US Government Printing Office; January 2007.

20. World Health Organization. Laboratory Biosafety Manual. 3rd ed. Geneva: World Health Organization; 2004.

21. Clinical and Laboratory Standards Institute. Protection of Laboratory Workers from Occupationally Acquired Infections: Approved Guideline —Third Edition. CLSI Document M29-A3. Wayne, PA: Clinical and Laboratory Standards Institute; 2005.Primus FJ, Kelley EA, Hansen HJ, Goldenberg DM. “Sandwich”-Type 22. Immunoassay of Carcinoembryonic Antigen in Patients Receiving Murine Monoclonal Antibodies for Diagnosis and Therapy. Clin Chem 1988; 34:261-4.

7

Schroff RW, Foon KA, Beatty SM, Oldham RK, Morgan AC Jr. 23. Human Anti-Murine Immunoglobulin Responses in Patients Receiving Monoclonal Antibody Therapy. Cancer Res 1985; 45:879-85.Boscato LM and Stuart MC. Heterophilic Antibodies; A Problem for 24. All Immunoassays. Clin Chem 1988; 34:27.National Committee for Clinical Laboratory Standards, Evaluation 25. of Precision Performance of Clinical Chemistry Devices - Approved Guideline. NCCLS Document EP5-A. Wayne, PA: NCCLS, 1999.Passing H, Bablok W. A New Biometrical Procedure for Testing the 26. Equality of Measurements from Two Different Analytical Methods. J Clin Chem. Clin Biochem. 1983;21:709-20.

ARCHITECT, AxSYM, MasterCheck and Chemiflex are trademarks of Abbott Laboratories in various jurisdictions.

Abbott IrelandDiagnostics DivisionLisnamuck, LongfordCo. LongfordIreland+353-43-3331000

Distributed by Abbott LaboratoriesAbbott Park, IL 60064 USAand ABBOTT 65205 Wiesbaden, Germany

June 2010© 2005, 2010 Abbott Laboratories

1

SYSTEM

TSH Controls

FINALIDAD DE USOLos ARCHITECT TSH Controls se utilizan para la verificación de la exactitud y la precisión del ARCHITECT i System en la determinación cuantitativa de la hormona tiroestimulante humana (TSH) en suero y plasma humanos. Si desea más información, consulte las instrucciones de uso del ensayo ARCHITECT correspondiente.

CONTENIDO3 frascos (8 ml cada uno) de ARCHITECT TSH Controls ( ,

, ), que contienen TSH (recombinante) preparada en tampón TRIS con estabilizantes proteínicos (bovinos). Conservante: azida sódica. Se pueden utilizar los siguientes intervalos de valores para las especificaciones de los controles en el sistema ARCHITECT:

Control

Concentración esperada / Intervalo de valores

µIU/ml

Concentración esperada/ Intervalo de valores mIU/l

0,1 0,065 - 0,135 0,1 0,065 - 0,135

6 3,9 - 8,1 6 3,9 - 8,1

30 19,5 - 40,5 30 19,5 - 40,5

Cada laboratorio debe establecer sus propios intervalos de concentración aceptables para cada lote de controles nuevo y para todas las concentraciones de los controles. Para ello, se debe analizar un mínimo de 20 replicados durante varios días (de 3 a 5 días). Entre las causas de variaciones que se pueden dar y que se deben incluir en este estudio para que sea representativo del funcionamiento futuro del sistema se incluyen:• Diversas calibraciones almacenadas• Diversos lotes de reactivos• Diversos lotes de calibradores• Diferentes módulos de procesamiento• Datos recogidos en diversos momentos del díaEstos resultados se deben utilizar en los procesos de control de calidad de su laboratorio.

ESTANDARIZACIÓNLos controles se fabrican mediante la adición de concentraciones conocidas de TSH humana recombinante para obtener una concentración esperada. Esta concentración asignada internamente se establece con valores de calibración que se correlacionan con el patrón 80/558 de la OMS para la TSH.

PRECAUCIONES•• Para uso en diagnóstico in vitro• Contiene azida sódica. En contacto con ácidos

libera gases muy tóxicos.• Elimínense los residuos del producto y sus recipientes con todas las

precauciones posibles.• Las fichas de datos de seguridad están disponibles en la página

web www.abbottdiagnostics.com o a través de la Asistencia Técnica de Abbott.

• Si desea más información sobre la eliminación correcta de los reactivos que contienen azida sódica, consulte el capítulo 8 del Manual de operaciones del sistema ARCHITECT.

ALMACENAMIENTO• Si se almacenan y se manejan según las instrucciones, los ARCHITECT

TSH Controls se mantienen estables hasta la fecha de caducidad.• No los utilice una vez transcurrida la fecha de caducidad.

•

ARCHITECT es una marca comercial de Abbott Laboratories en varios países.


Distribuido por Abbott LaboratoriesAbbott Park, IL 60064 USAy ABBOTT 65205 Wiesbaden, Germany

Noviembre 2012 © 2004, 2012 Abbott Laboratories

Símbolos utilizados

Código GTIN, número mundial de identificación de artículo

Producto de Irlanda

Información de interés sólo para EE. UU.

Consulte las modificaciones marcadasRevisado en noviembre de 2012

esTSH

7K62-10

C7K623G3-3941/R03

1

system

TSH Calibrators

FINALIDAD DE USOLos ARCHITECT TSH Calibrators se usan para la calibración del ARCHITECT i System en la determinación cuantitativa de la hormona tiroestimulante humana (TSH) en suero y plasma humanos. Si desea más información, consulte las instrucciones de uso específicas del ensayo ARCHITECT correspondiente.

CONTENIDO2 frascos (4 ml cada uno) de ARCHITECT TSH Calibrators. El calibrador 1 ( ) contiene tampón TRIS con estabilizantes proteínicos (bovinos). El calibrator 2 ( ) contiene TSH (recombinante) en tampón TRIS con estabilizantes proteínicos (bovinos). Conservante: azida sódica.Los calibradores presentan las concentraciones siguientes:

CalibradorConcentración de TSH

(μIU/ml) (mIU/l)0 0

40 40

ESTANDARIZACIÓNLos calibradores se elaboran añadiendo una concentración conocida de TSH humana recombinante para obtener una concentración deseada. La concentración se ajusta al patrón 80/558 para TSH de la Organización Mundial de la Salud (OMS).

PRECAUCIONES•• Para uso en diagnóstico in vitro• Contiene azida sódica. En contacto con ácidos

libera gases muy tóxicos.• Elimínense los residuos del producto y sus recipientes con todas las

precauciones posibles.• Las fichas de datos de seguridad están disponibles en la página

web www.abbottdiagnostics.com o a través de la Asistencia Técnica de Abbott.

• Si desea más información sobre la eliminación correcta de los reactivos que contienen azida sódica, consulte el capítulo 8 del Manual de operaciones del sistema ARCHITECT.

ALMACENAMIENTO• Si se almacenan y se manejan según las instrucciones, los ARCHITECT

TSH Calibrators se mantienen estables hasta la fecha de caducidad. • No los utilice una vez transcurrida la fecha de caducidad.

•

ARCHITECT es una marca comercial de Abbott Laboratories en varios países.


Distribuido por Abbott LaboratoriesAbbott Park, IL 60064 USAy ABBOTT 65205 Wiesbaden, Germany

Noviembre 2012 © 2004, 2012 Abbott Laboratories

Símbolos utilizados

Código GTIN, número mundial de identificación de artículo

Producto de Irlanda

Información de interés sólo para EE. UU.

Consulte las modificaciones marcadasRevisado en noviembre de 2012

esTSH

7K62-01

S7K623G3-3930/R03

1

system

en25-OH Vitamin D

3L52

49-8941/R02

25-OH Vitamin D

Customer Service: Contact your local representative or find country specific contact information on www.abbottdiagnostics.com

Package insert instructions must be carefully followed. Reliability of assay results cannot be guaranteed if there are any deviations from the instructions in this package insert.

Key to symbols used

List Number

In Vitro Diagnostic Medical Device

Lot Number

Expiration Date

Serial Number

Store at 2-8°C


Manufacturer

Control Number

Reaction Vessels

Sample Cups

Septum

Replacement Caps

Warning: May cause an allergic reaction

Contains sodium azide. Contact with acids liberates very toxic gas.

Contains: Methanol

Assay CD-ROM - WW (excluding US) - Addition E

Assay CD-ROM - WW (excluding US) - Addition E

See REAGENTS section for a full explanation of symbols used in reagent component naming.

Read Highlighted ChangesRevised June 2011

2

NAMEARCHITECT 25-OH Vitamin D

INTENDED USEThe ARCHITECT 25-OH Vitamin D assay is a chemiluminescent microparticle immunoassay (CMIA) for the quantitative determination of 25-hydroxyvitamin D (25-OH vitamin D) in human serum and plasma. The ARCHITECT 25-OH Vitamin D assay is to be used as an aid in the assessment of vitamin D sufficiency.

SUMMARY AND EXPLANATION OF TESTVitamin D is a fat-soluble steroid prohormone mainly produced photochemically in the skin from 7-dehydrocholesterol.Two forms of vitamin D are biologically relevant – vitamin D3 (Cholecalciferol) and vitamin D2 (Ergocalciferol). Both vitamins D3 and D2 can be absorbed from food, with vitamin D2 being an artificial source, but only an estimated 10-20% of vitamin D is supplied through nutritional intake.1 Vitamins D3 and D2 can be found in vitamin supplements. Vitamin D is converted to the active hormone 1,25-(OH)2-vitamin D (Calcitriol) through two hydroxylation reactions. The first hydroxylation converts vitamin D into 25-OH vitamin D and occurs in the liver. The second hydroxylation converts 25-OH vitamin D into the biologically active 1,25-(OH)2-vitamin D and occurs in the kidneys as well as in many other cells of the body. Most cells express the vitamin D receptor and about 3% of the human genome is directly or indirectly regulated by the vitamin D endocrine system.1

The major storage form of vitamin D is 25-OH vitamin D and is present in the blood at up to 1,000 fold higher concentration compared to the active 1,25-(OH)2-vitamin D. 25-OH vitamin D has a half-life of 2-3 weeks vs. 4 hours for 1,25-(OH)2-vitamin D. Therefore, 25-OH vitamin D is the analyte of choice for determination of the vitamin D status.2,3 Epidemiological studies have shown a high global prevalence of vitamin D insufficiency and deficiency.4 The measurement of vitamin D status provides opportunities for preventive and therapeutic interventions.5,6,7

Vitamin D deficiency is a cause of secondary hyperparathyroidism and diseases resulting in impaired bone metabolism (like rickets, osteoporosis, osteomalacia).2,8,9 Reduced 25-OH vitamin D concentrations in blood (vitamin D insufficiency) have been associated with an increasing risk of many chronic diseases, including common cancers, autoimmune or infectious diseases or cardiovascular problems.1,2,6,8,10-12

BIOLOGICAL PRINCIPLES OF THE PROCEDUREThe ARCHITECT 25-OH Vitamin D assay is a delayed one-step immunoassay including a sample pre-treatment for the quantitative determination of vitamin D in human serum and plasma using CMIA technology with flexible assay protocols, referred to as Chemiflex.Sample and pre-treatment reagent are combined. An aliquot of the pre-treated sample is combined with assay diluent and paramagnetic anti-vitamin D coated microparticles to create a reaction mixture. Vitamin D present in the sample binds to anti-vitamin D coated microparticles. After incubation a biotinylated vitamin D anti-Biotin acridinium-labeled conjugate complex is added to the reaction mixture and binds to unoccupied binding sites of the anti-vitamin D coated microparticles. After washing, pre-trigger and trigger solutions are added to the reaction mixture. The resulting chemiluminescent reaction is measured as relative light units (RLUs). An indirect relationship exists between the amount of vitamin D in the sample and the RLUs detected by the ARCHITECT i System optics. For additional information on system and assay technology, refer to the ARCHITECT System Operations Manual, Section 3.

REAGENTSReagent Kit, 100 Tests/500 TestsNOTE: Some kit sizes are not available in all countries or for use on all ARCHITECT i Systems. Please contact your local distributor.ARCHITECT Reagent Kit (3L52)• 1 Bottle (13.3 mL per 100-test bottle/27.0 mL per

500-test bottle) Anti-human vitamin D IgG (sheep, polyclonal) coated microparticles in TRIS buffer. Minimum concentration: 0.05% solids. Preservatives: ProClin 300, ProClin 950.

• 1 Bottle (5.9 mL per 100-test bottle/26.3 mL per 500-test bottle) biotinylated vitamin D anti-biotin IgG (mouse, monoclonal) acridinium-labeled conjugate complex in BIS-TRIS HCl buffer with protein stabilizers (bovine gamma globulin) and detergent. Minimum concentration: 1.2 μg/mL anti-biotin IgG and 0.1 μg/mL vitamin-D-biotin. Preservative: sodium azide.

• 1 Bottle (4.9 mL per 100-test bottle/21.2 mL per 500-test bottle) Assay Diluent containing acetic acid buffer with EDTA. Preservatives: ProClin 300, ProClin 950.

• 1 Bottle (10.0 mL per 100-test bottle/50.9 mL per 500-test bottle) Pre-Treatment 1 containing triethanolamine methanol buffer and 8-anilino-1-naphtalensulfonic acid (ANSA).

• 1 Bottle (5.9 mL per 100-test bottle/26.3 mL per 500-test bottle) Pre-Treatment 2 containing triethanolamine methanol buffer and 8-anilino-1-naphtalensulfonic acid (ANSA).

Other ReagentsARCHITECT i Pre-Trigger Solution• Pre-trigger solution containing 1.32% (w/v)

hydrogen peroxide.ARCHITECT i Trigger Solution• Trigger solution containing 0.35N sodium

hydroxide.ARCHITECT i Wash Buffer• Wash buffer containing phosphate buffered saline

solution. Preservatives: antimicrobial agents.

WARNINGS AND PRECAUTIONS•• For In Vitro Diagnostic Use.Package insert instructions must be carefully followed. Reliability of assay results cannot be guaranteed if there are any deviations from the instructions in this package insert.

Safety Precautions• CAUTION: This product requires the handling of human specimens.

It is recommended that all human-sourced materials be considered potentially infectious and handled in accordance with the OSHA Standard on Bloodborne Pathogens.13 Biosafety Level 214 or other appropriate biosafety practices15,16 should be used for materials that contain or are suspected of containing infectious agents.

• This product contains sodium azide; for a specific listing, refer to the REAGENTS section. Contact with acids liberates very toxic gas. This material and its container must be disposed of in a safe way.

• The following warnings and precautions apply to these components:• Microparticles• Assay Diluent

WARNING: Contains methylisothiazolones.H317 May cause an allergic skin reaction.PreventionP261 Avoid breathing mist/vapours/spray.P272 Contaminated work clothing should not be

allowed out of the workplace.P280 Wear protective gloves/protective clothing/eye

protection.ResponseP302+P352 IF ON SKIN: Wash with plenty of water.P333+P313 If skin irritation or rash occurs: Get medical

advice/attention.P363 Wash contaminated clothing before reuse.

This material and its container must be disposed of in a safe way.

3

• The following warnings and precautions apply to these components:• Pre-Treatment 1• Pre-Treatment 2

DANGER: Contains methanol.H226 Flammable liquid and vapour.H332 Harmful if inhaled.H370 Causes damage to organs.Prevention

P210 Keep away from heat/sparks/open flames/hot surfaces. No smoking.

P243 Take precautionary measures against static discharge.

P233 Keep container tightly closed.P271 Use only in a well-ventilated area.P260 Do not breathe mist/vapours/spray.P280 Wear protective gloves/protective clothing/eye

protection.P264 Wash hands thoroughly after handling.ResponseP304+P340 IF INHALED: Remove victim to fresh air and

keep at rest in a position comfortable for breathing.

P303+P361+ P353

IF ON SKIN (or hair): Remove/Take off immediately all contaminated clothing. Rinse skin with water/shower.

P307+P311 IF EXPOSED: Call a POISON CENTER or doctor/physician.

StorageP403+P235 Store in a well-ventilated place. Keep cool.P405 Store locked up.

This material and its container must be disposed of in a safe way.

• For information on the safe disposal of sodium azide and a detailed discussion of safety precautions during system operation, refer to the ARCHITECT System Operations Manual, Section 8.

Handling Precautions• Do not use reagent kits beyond the expiration date.• Do not pool reagents within a kit or between reagent kits.• The conjugate is temperature sensitive, minimize room temperature

exposure of conjugate bottle.• Before loading the ARCHITECT 25-OH Vitamin D Reagent Kit on

the system for the first time, the microparticle bottle requires mixing to resuspend microparticles that have settled during shipment. For microparticle mixing instructions, refer to the PROCEDURE, Assay Procedure section of this package insert.

• Septums MUST be used to prevent reagent evaporation and contamination and to ensure reagent integrity. Reliability of assay results cannot be guaranteed if septums are not used according to the instructions in this package insert.

• To avoid contamination, wear clean gloves when placing a septum on an uncapped reagent bottle.• Once a septum has been placed on the reagent bottle, do not

invert the bottle as this will result in reagent leakage and may compromise assay results.

• Over time, residual liquids may dry on the septum surface. These are typically dried salts, which have no effect on assay efficacy.

• For a detailed discussion of handling precautions during system operation, refer to the ARCHITECT System Operations Manual, Section 7.

Storage Instructions

• The ARCHITECT 25-OH Vitamin D Reagent Kit must be stored at 2-8°C in an upright position and may be used immediately after removal from 2-8°C storage.

• When stored and handled as directed, reagents are stable until the expiration date.

• The conjugate is temperature sensitive, minimize room temperature exposure of conjugate bottle.

• The ARCHITECT Reagent Kit may be stored on board the ARCHITECT i System for a maximum of 14 days.

• After these maximum storage times, the reagent kit must be discarded. For information on tracking onboard time, refer to the ARCHITECT System Operations Manual, Section 5.

• Reagents may be stored on or off the ARCHITECT i System. If reagents are removed from the system, store them at 2-8°C (with septums and replacement caps) in an upright position. For reagents stored off the system, it is recommended that they be stored in their original trays and boxes to ensure they remain upright. If the microparticle bottle does not remain upright (with a septum installed) while in refrigerated storage off the system, the reagent kit must be discarded. For information on unloading reagents, refer to the ARCHITECT System Operations Manual, Section 5.

Indications of Reagent DeteriorationWhen a control value is out of the specified range, it may indicate deterioration of the reagents or errors in technique. Associated test results are invalid and samples must be retested. Assay recalibration may be necessary. For troubleshooting information, refer to the ARCHITECT System Operations Manual, Section 10.

INSTRUMENT PROCEDURE• The ARCHITECT 25-OH Vitamin D assay requires:

• ARCHITECT i System e-Assay CD-ROM found on www.abbottdiagnostics.com under Support/Technical Library/

Assay Files or• ARCHITECT i System Assay CD-ROM - WW (excluding US) -

Addition E.• The ARCHITECT 25-OH Vitamin D assay file must be installed on

the ARCHITECT i System before performing the assay. For detailed information on assay file installation and on viewing and editing assay parameters, refer to the ARCHITECT System Operations Manual, Section 2.

• For information on printing assay parameters, refer to the ARCHITECT System Operations Manual, Section 5.

• For a detailed description of system procedures, refer to the ARCHITECT System Operations Manual.

• The default result unit for the ARCHITECT 25-OH Vitamin D assay is ng/mL. An alternate result unit, nmol/L, may be selected for reporting results by editing assay parameter “Result concentration units” to nmol/L. The conversion factor used by the system is 2.5.

SPECIMEN COLLECTION AND PREPARATION FOR ANALYSISSpecimen TypesThe specimen collection tubes listed below were verified to be used with the ARCHITECT 25-OH Vitamin D assay. Other specimen collection tubes have not been tested with this assay.• Human serum (including serum collected in serum separator tubes)• Human plasma collected in:

• Potassium-EDTA• Lithium heparin (powder or gel)• Sodium heparin

• Human plasma collected in sodium citrate tube types showed a mean bias of -13% versus serum.

• Other liquid anticoagulants may also have a dilution effect resulting in lower concentrations for individual patient specimens.

• The ARCHITECT i System does not provide the capability to verify specimen type. It is the responsibility of the operator to verify that the correct specimen types are used in the ARCHITECT 25-OH Vitamin D assay.

4

Specimen Conditions• Do not use specimens with the following conditions:

• heat-inactivated• pooled• grossly hemolyzed (> 500 mg/dL hemoglobin)• obvious microbial contamination• cadaver specimens or any other body fluids

• For accurate results, serum and plasma specimens should be free of fibrin, red blood cells, and other particulate matter. Serum specimens from patients receiving anticoagulant or thrombolytic therapy may contain fibrin due to incomplete clot formation.

• Use caution when handling patient specimens to prevent cross contamination. Use of disposable pipettes or pipette tips is recommended.

• For optimal results, inspect all specimens for bubbles. Remove bubbles with an applicator stick before analysis. Use a new applicator stick for each specimen to prevent cross contamination.

Preparation for Analysis• Follow the tube manufacturer’s processing instructions for serum

and plasma collection tubes. Gravity separation is not sufficient for specimen preparation.

• Mix thawed specimens thoroughly by low speed vortexing or by inverting 10 times. Visually inspect the specimens. If layering or stratification is observed, continue mixing until specimens are visibly homogeneous.

• To ensure consistency in results, specimens must be transferred to a centrifuge tube and centrifuged at ≥ 10000 RCF (Relative Centrifugal Force) for 10 minutes before testing if• they contain fibrin, red blood cells, or other particulate matter,• they require repeat testing, or• they were frozen and thawed.

Transfer clarified specimen to a sample cup or secondary tube for testing.

• Centrifuged specimens with a lipid layer on the top must be transferred to a sample cup or secondary tube. Care must be taken to transfer only the clarified specimen without the lipemic material.

Storage• Specimens may be stored on or off the clot or red blood cells for:

• up to 12 days at 2-8°C or• up to 72 hours at 15-30°C

• If testing will be delayed more than 12 days, remove serum or plasma from the clot or red blood cells and store frozen at -20°C or colder.

• Specimens stored frozen for 3 months showed no performance difference. Avoid more than 4 freeze/thaw cycles.

Shipping• Before shipping specimens, it is recommended that specimens be

removed from the clot, red blood cells, or separator gel.• When shipping specimens, package and label specimens in compliance

with applicable state, federal, and international regulations covering the transport of clinical specimens and infectious substances.

• Specimens may be shipped: ambient, on wet ice, on dry ice. Do not exceed the storage time limitations listed above.

PROCEDUREMaterials Provided• 3L52 ARCHITECT 25-OH Vitamin D Reagent Kit

Materials Required but not Provided• ARCHITECT i System• 1L66 ARCHITECT i System

Assay CD-ROM - WW (excluding US) - Addition E , Version 9.0 or higher or

• ARCHITECT i System e-Assay CD-ROM found on www.abbottdiagnostics.com under Support/Technical Library/Assay

Files.• 3L52-01 ARCHITECT 25-OH Vitamin D Calibrators• 3L52-10 ARCHITECT 25-OH Vitamin D Controls• ARCHITECT i • ARCHITECT i

• ARCHITECT i • ARCHITECT i • ARCHITECT i • ARCHITECT i • ARCHITECT i • Pipettes or pipette tips (optional) to deliver the volumes specified on

the patient or control order screen.For information on materials required for maintenance procedures, refer to the ARCHITECT System Operations Manual, Section 9.

Assay Procedure• Before loading the ARCHITECT 25-OH Vitamin D Reagent Kit on the

system for the first time, the microparticle bottle requires mixing to resuspend microparticles that have settled during shipment. After the first time the microparticles have been loaded, no further mixing is required.• Invert the microparticle bottle 30 times. • Visually inspect the bottle to ensure microparticles are

resuspended. If microparticles remain adhered to the bottle, continue inverting the bottle until the microparticles have been completely resuspended.

• If the microparticles do not resuspend, DO NOT USE. Contact your local Abbott representative.

• Once the microparticles have been resuspended, place a septum on the bottle. For instructions on placing septums on bottles refer to the Handling Precautions section of this package insert.

• Load the ARCHITECT 25-OH Vitamin D Reagent Kit on the ARCHITECT i System.• Verify that all necessary assay reagents are present.• Ensure that septums are present on all reagent bottles.

• Order calibration, if necessary.• For information on ordering calibrations, refer to the ARCHITECT

System Operations Manual, Section 6.• Order tests.

• For information on ordering patient specimens and controls and for general operating procedures, refer to the ARCHITECT System Operations Manual, Section 5.

• The minimum sample cup volume is calculated by the system and is printed on the Orderlist report. No more than 10 replicates may be sampled from the same sample cup. To minimize the effects of evaporation, verify adequate sample cup volume is present before running the test.• Priority: 60 μL for the first ARCHITECT 25-OH Vitamin D test plus

10 μL for each additional ARCHITECT 25-OH Vitamin D test from the same sample cup.

• ≤ 3 hours on board: 150 µl for the first ARCHITECT 25-OH Vitamin D test plus 10 µl for each additional ARCHITECT 25-OH Vitamin D test from the same sample cup.

• > 3 hours on board: additional sample volume is required. For information on sample evaporation and volumes, refer to the ARCHITECT System Operations Manual, Section 5.

• If using primary or aliquot tubes, use the sample gauge to ensure sufficient patient specimen is present.

• Prepare calibrators and controls.• Mix ARCHITECT 25-OH Vitamin D Calibrators and Controls by

gentle inversion before use.• To obtain the recommended volume requirements for the

ARCHITECT 25-OH Vitamin D Calibrators and Controls, hold the bottles vertically and dispense 5 drops of each calibrator or 5 drops of each control into each respective sample cup.

• Load samples.• For information on loading samples, refer to the ARCHITECT

System Operations Manual, Section 5.• Press RUN.• For additional information on principles of operation, refer to the

ARCHITECT Operations Manual, Section 3.• For optimal performance, it is important to perform routine

maintenance as described in the ARCHITECT System Operations Manual, Section 9. Perform maintenance more frequently when required by laboratory procedures.

5

Specimen Dilution ProceduresSpecimens with a 25-OH vitamin D concentration > 160.0 ng/mL (> 400.0 nmol/L) will be flagged as “>160.0 ng/mL” (“>400.0 nmol/L”) and may be diluted with a specimen with a low 25-OH vitamin D concentration ≤ 10.7 ng/mL (≤ 26.8 nmol/L).Manual dilutions should be performed as follows: Add 100 μL of the patient specimen to 100 μL of a specimen with a low 25-OH vitamin D concentration (i.e. 1:2 dilution).Concentration (Conc.) calculations are performed as follows:Conc. (high specimen) = Conc. (1:2 diluted specimen) x 2 - Conc. (low specimen)

Calibration• The ARCHITECT 25-OH Vitamin D assay requires a 7-days

recalibration interval.• To perform an ARCHITECT 25-OH Vitamin D calibration, test

calibrators A, B, C, D, E, and F in replicates of two. A single sample of each 25-OH vitamin D control level must be tested to evaluate the assay calibration. Ensure that assay control values are within the concentration ranges specified in the control package insert. Calibrators should be priority loaded.

• Calibration Range: 0.0 to 160.0 ng/mL (0.0 to 400.0 nmol/L).• For detailed information on how to perform an assay calibration, refer

to the ARCHITECT System Operations Manual, Section 6.

QUALITY CONTROL PROCEDURESThe recommended control requirement for the ARCHITECT 25-OH Vitamin D assay is that a single sample of each control be tested once every 24 hours each day of use. If your laboratory quality control procedures require more frequent use of controls to verify test results, follow those procedures.Additional controls may be tested in conformance with local, state, and/or federal regulations or accreditation requirements and your laboratory´s quality control procedure.Each laboratory should establish control ranges to monitor the acceptable performance of the assay. If a control is out of its specified range, the associated test results are invalid and samples must be retested. Recalibration may be indicated.

Verification of Assay ClaimsFor protocols to verify package insert claims, refer to the ARCHITECT System Operations Manual, Appendix B. The ARCHITECT 25-OH Vitamin D assay belongs to method group 2.

RESULTSThe ARCHITECT 25-OH Vitamin D assay uses a 4 Parameter Logistic Curve fit (4PLC, Y-weighted) data reduction method to generate a calibration curve.

CalculationThe ARCHITECT i System calculates the Calibrator A through F mean chemiluminescent signal from two Calibrator A through F replicates, generates a calibration curve and stores the result. The default result unit for the ARCHITECT 25-OH Vitamin D assay is ng/mL.

FlagsSome results may contain information in the Flags field. For a description of the flags that may appear in this field, refer to the ARCHITECT System Operations Manual, Section 5.

Measuring Interval (Reportable Range)The measuring interval of the ARCHITECT 25-OH Vitamin D assay is 8.0 ng/mL to 160.0 ng/mL (20.0 nmol/L to 400.0 nmol/L).

LIMITATIONS OF THE PROCEDURE• If the vitamin D results are inconsistent with clinical evidence,

additional testing is suggested to confirm the result.• For diagnostic purposes, results should be used in conjunction with

other data; e.g., symptoms, results of other tests, clinical impressions, etc.

• Heterophilic antibodies in human serum can react with reagent immunoglobulins, interfering with in vitro immunoassays.17 Patients routinely exposed to animals or to animal serum products can be prone to this interference and anomalous values may be observed. Additional information may be required for diagnosis.

• Specimens from patients who have received preparations of mouse monoclonal antibodies for diagnosis or therapy may contain human anti-mouse antibodies (HAMA).18,19 Specimens containing HAMA may produce anomalous values when tested with assay kits that employ mouse monoclonal antibodies.18

EXPECTED VALUESIt is recommended that each laboratory establish its own reference range, which may be unique to the population it serves depending upon geographical, season, patient, dietary, or environmental factors.A reference range study was conducted based on guidance from the Clinical Laboratory and Standards Institute (CLSI), Protocol C28-A320. Serum specimens of apparently healthy individuals from the Netherlands, collected during summertime (June to August) and wintertime (March to May, November and December) were evaluated in replicates of one using the ARCHITECT 25-OH Vitamin D assay. A medical questionnaire has been used to select the reference sample group based on the following inclusion criteria: age between 17 and 60 years, normal nutrition habits (no diet), no food supplements, no bone disease or rheumatism, no recent sunny vacation or sunbath, no current hospital treatment, no chronic disease (especially diabetes, renal failure, high cholesterol). In addition, individuals have been excluded when showing results out of the expected values for i PTH (15.0 – 68.3 pg/mL) and calcium (2.10 – 2.55 mmol/L). The observed values are summarized in the following table.*

All

SeasonGender Summer-

timeWinter-

time UnitFemale Malen 360 178 182 120 240

2.5th Percentile 9.5 9.4 9.4 15.7 8.8 ng/mL97.5th Percentile 55.5 59.1 52.4 60.3 46.3 ng/mL

* Representative data; results in individual laboratories and in different geographical areas may vary from these data.

There is currently debate over the recommended target range of vitamin D in serum, but an expert panel recently suggested a target range of at least 30 - 40 ng/mL.21

SPECIFIC PERFORMANCE CHARACTERISTICSData in the section SPECIFIC PERFORMANCE CHARACTERISTICS were generated using the ARCHITECT i2000SR System.Assay results obtained in individual laboratories may vary from data presented.

PrecisionThe ARCHITECT 25-OH Vitamin D assay is designed to have an imprecision of ≤ 10% Within Laboratory (total) CV.A study was performed with the ARCHITECT 25-OH Vitamin D assay based on guidance from the National Committee for Clinical Laboratory Standards (NCCLS) Protocol EP5-A222. Six samples consisting of 3 ARCHITECT 25-OH Vitamin D Controls and 3 serum based panels were assayed, using two lots of reagents, on one instrument, in replicates of two at two separate times per day for 20 days. Data from this study are summarized in the following table.

SampleReagent

Lot n

Mean Conc.

(ng/mL)Within Run

Within Laboratory (total)

SD %CV SD %CVLow Control

1 80 19.0 0.709 3.7 0.712 3.82 80 19.5 0.589 3.0 0.889 4.6

Medium Control

1 80 38.5 0.873 2.3 1.142 3.02 80 38.0 0.879 2.3 1.062 2.8

High Control

1 80 78.4 1.470 1.9 2.201 2.82 80 76.3 1.485 1.9 2.034 2.7

Serum Panel 1

1 80 23.0 0.714 3.1 0.912 4.02 80 22.4 0.548 2.4 0.780 3.5

Serum Panel 2

1 80 42.5 1.095 2.6 1.346 3.22 80 40.1 0.668 1.7 1.274 3.2

Serum Panel 3

1 80 75.4 1.088 1.4 2.064 2.72 80 71.3 1.242 1.7 1.869 2.6

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Linear RangeBased on guidance from the NCCLS Protocol EP6-A23 a study was performed to establish the linear range of the ARCHITECT 25-OH Vitamin D assay. Four sample pairs were prepared with 11 dilutions for each pair by mixing a high 25-OH vitamin D sample (in the range from 78.4 ng/mL to 165.5 ng/ mL) in specific ratios with a low 25-OH vitamin D sample (in the range from 0.2 ng/mL to 10.7 ng/mL) and tested using the ARCHITECT 25-OH Vitamin D assay. Using an absolute deviation from linearity of ≤ 20% a linear range of 9.4 ng/mL to 165.5 ng/mL was established for the ARCHITECT 25-OH Vitamin D assay.

Measuring IntervalMeasuring interval is defined as the range of values in ng/mL which meets the limits of acceptable performance for both imprecision and bias for an undiluted sample. For the verification studies described in this package insert, the range was 8.0 ng/mL to 160.0 ng/mL (20.0 nmol/L to 400.0 nmol/L).

SensitivityThe ARCHITECT 25-OH Vitamin D assay is designed to have a Limit of Blank (LoB) of ≤ 4.0 ng/mL, a Limit of Detection (LoD) of ≤ 10.0 ng/mL and a Limit of Quantitation (LoQ) of ≤ 20 ng/mL.Based on guidance from the NCCLS Protocol EP17-A24 a study was performed with 4 zero-level samples (Calibrator A) and 8 samples with 25-OH vitamin D concentrations ranging from 3.4 ng/mL to 9.5 ng/ mL. These samples were tested in 5 separate runs over a minimum of 5 days using two reagent lots and two instruments. In the above described study the LoB was 1.9 ng/mL, the LoD was 3.1 ng/ mL and the LoQ was 8.0 ng/mL.

SpecificityThe specificity of the ARCHITECT 25-OH Vitamin D assay was assessed by testing the Cross-Reactants listed in the table below.A study was performed with the ARCHITECT 25-OH Vitamin D assay based on guidance from the CLSI Protocol EP7-A225. Aliquots of ARCHITECT 25-OH Vitamin D Calibrator A were supplemented with potential Cross-Reactants at the concentrations listed and tested for 25-OH vitamin D. Data from this study are summarized in the following table.

Cross-ReactantConcentration

ng/mL% Cross-

Reactivitya

25-OH vitamin D3 100 105Vitamin-D2 (Ergocalciferol) 1000 0.1Vitamin-D3 (Cholecalciferol) 1000 0.324,25-(OH)2-vitamin D3 20 1121,25-(OH)2-vitamin D3 100 12.63-epi 25-OH vitamin D3 100 2.7Paricalcitol (Zemplar) 24 0.4

a % Cross-Reactivity =

Mean Value spiked (ng/mL) - Mean Value non spiked (ng/mL) x 100

Concentration of Cross-Reactant (ng/mL)

The 25-OH vitamin D2 % Cross-Reactivity* has been determined using endogenous (non-spiked) serum samples. The samples were analyzed with a chromatographic method (LC-MS/MS) in order to determine 25-OH vitamin D2 and 25-OH vitamin D3 concentration. Samples have been included in the testing that showed a 25-OH vitamin D2 to 25-OH vitamin D3 ratio of ≥ 10 and 25-OH vitamin D3 concentration < 5.0 ng/mL.

* D2 % Cross-Reactivity =

25-OH vitamin Total (Arch) - 25-OH vitamin D3 (LC – MS/MS) x 10025-OH vitamin D2 (LC – MS/MS)

The mean D2 % Cross-Reactivity for ARCHITECT 25-OH Vitamin D was 82.

Interference Potential interference in the ARCHITECT 25-OH Vitamin D assay from hemoglobin, bilirubin, triglycerides, protein, rheumatoid factor, HAMA and red blood cells is designed to be ≤ 10%.

Interference was demonstrated by a study based on guidance from the CLSI Protocol EP7-A2. Data from this study are summarized in the following table.

Potentially Interfering Substance Concentration

Range of % Recoverya

Hemoglobinb 200 mg/dL 91 - 98Bilirubin 20 mg/dL 100 - 104Triglycerides 5000 mg/dL 94 - 103Protein (Human Albumin) 12 g/dL 90 - 102Rheumatoid Factorc 400 IU/mL 106 - 109HAMA 1000 ng/mL 96 - 99Red blood cells 0.4 %(v/v) 99 - 103

a % Recovery = Mean Observed Value (ng/mL) x 100

Mean Control Value (ng/mL)b For hemoglobin with concentrations between 200 mg/dL and

500 mg/dL the % Recovery ranged from 90% to 62%.c For rheumatoid factors with concentrations between 400 IU/mL and

800 IU/mL the % Recovery ranged from 107% to 118%.

Method ComparisonThe ARCHITECT 25-OH Vitamin D assay is designed to have a correlation coefficient of ≥ 0.80 for serum samples when evaluated against the DiaSorin LIAISON 25-OH Vitamin D Total. A study was performed with the ARCHITECT 25-OH Vitamin D assay, where regression analysis was performed using the Passing-Bablok26 method. Data from this study are summarized in the following table and graph.*

Regression Method n Slope Intercept

Correlation Coefficient

Passing-Babloka 108 1.02 -0.54 0.94a A linear regression method with no special assumptions regarding the

distribution of the samples and measurement errors.* Representative data; variables such as differences in sampling size and

sample population may impact the correlation of the assay, therefore, results in individual laboratories may vary from these data.

In this evaluation, serum specimen concentrations ranged from 8.2 ng/ mL to 70.5 ng/mL with the ARCHITECT 25-OH Vitamin D assay and from 5.1 ng/mL to 63.2 ng/mL with the DiaSorin LIAISON 25-OH Vitamin D Total. The specimens included in the study are sourced from a clinical research organization.

ARCHITECT 25-OH Vitamin D vs.

DiaSorin LIAISON 25-OH Vitamin D Total (Passing-Bablok)

DiaSorin LIAISON 25-OH Vitamin D Total (ng/mL)

7

The same specimens have been measured with a chromatographic method (LC-MS/MS) and evaluated against the ARCHITECT 25-OH Vitamin D assay. The regression analysis was performed using the Passing-Bablok method. Data from this study are summarized in the following table and graph.*

Regression Method n Slope Intercept

Correlation Coefficient

Passing-Babloka 107 1.01 - 1.45 0.90a A linear regression method with no special assumptions regarding the

distribution of the samples and measurement errors.* Representative data; variables such as differences in sampling size

and sample population may impact the correlation of the assay, therefore, results in individual laboratories may vary from these data. Additionally, LC-MS/MS methodology variables, such as extraction method, ionization method, type of chromatographic separation and type of internal standard may impact the LC-MS/MS results; therefore results in individual laboratories may vary from these data.27

In this evaluation, serum specimen concentrations ranged from 8.2 ng/mL to 70.5 ng/mL with the ARCHITECT 25-OH Vitamin D assay and from 7.5 ng/mL to 64.4 ng/mL with the LC-MS/MS test method.

ARCHITECT 25-OH Vitamin D vs.

LC-MS/MS 25-OH Vitamin D (Passing-Bablok)

LC-MS/MS 25-OH Vitamin D (ng/mL)

BIBLIOGRAPHY1. Pilz S, Vitamin D status and arterial hypertension: a systematic

review. Nat. Rev. Cardiol. 2009;6:621-30.2. Souberbielle JC, Evaluating vitamin D status. Implications for

preventing and managing osteoporosis and other chronic diseases. Joint Bone Spine 2006;73:249-53.

3. Cavalier E, Vitamin D: current status and perspectives. Clin Chem Lab Med 2009;47(2):120-27.

4. Peterlik M, Vitamin D and calcium insufficiency-related chronic diseases: an emerging world-wide public health problem. Int. J. Environ. Res. Public Health 2009;6:2585-607.

5. Grant WB, Estimated benefit of increased vitamin D status in reducing the economic burden of disease in western Europe. Prog. Biophys. Mol. Biol. 2009;99:104-13.

6. Holick MF, Vitamin D deficiency. N. Engl. J. Med. 2007;357:266-81.7. Autier P, Vitamin D supplementation and total mortality: A meta-

analysis of randomized controlled trials. Arch Intern Med. 2007; 167(16):1730-7.

8. Bischoff-Ferrari HA, Estimation of optimal serum concentrations of 25-hydroxyvitamin D for multiple health outcomes. Am J Clin Nutr 2006;84:18-28.

9. Steingrimsdottir L, Relationship between serum parathyroid hormone Levels, vitamin D sufficiency, and calcium intake. JAMA 2005; 294(18):2336-41.

10. Grant WB, Current impediments to acceptance of the ultraviolet-B-vitamin D-cancer hypothesis. Anticancer Res. 2009;29:3597-604.

11. Pilz S, Low vitamin D levels predict stroke in patients referred to coronary angiography. Stroke 2008;9:2611-13.

12. Bischoff-Ferrari HA, Fall prevention with supplemental and active forms of vitamin D: a meta-analysis of randomised controlled trials. BMJ 2009;339:b3692.

13. US Department of Labor, Occupational Safety and Health Administration, 29 CFR Part 1910.1030, Bloodborne pathogens.

14. US Department of Health and Human Services. Biosafety in microbiological and biomedical laboratories. 5th ed. Washington, DC: US Government Printing Office; January 2007.

15. World Health Organization. Laboratory Biosafety Manual. 3rd ed. Geneva: World Health Organization; 2004.

16. Clinical and Laboratory Standards Institute. Protection of Laboratory Workers from Occupationally Acquired Infections: Approved Guideline—Third Edition. CLSI Document M29-A3. Wayne, PA: Clinical and Laboratory Standards Institute; 2005.

17. Boscato, LM and Stuart, MC. Heterophilic antibodies; A problem for all immunoassays. Clin Chem 1988;34:27.

18. Primus FJ, Kelley EA, Hansen HJ, et al. “Sandwich”-type immunoassay of carcinoembryonic antigen in patients receiving murine monoclonal antibodies for diagnosis and therapy. Clin Chem 1988;34:261-4.

19. Schroff RW, Foon KA, Beatty SM, et al. Human anti-murine immunoglobulin responses in patients receiving monoclonal antibody therapy. Cancer Res 1985;45:879-85.

20. Clinical and Laboratory Standards Institute. Defining, Establishing, and Verifying Reference Intervals in the Clinical Laboratory: Approved Guideline - Third Edition. CLSI Document C28-A3. Wayne, PA: Clinical and Laboratory Standards Institute, 2008.

21. Souberbielle JC, Vitamin D and musculoskeletal health, cardiovascular disease, autoimmunity and cancer: Recommendations for clinical practice. Autoimmun Rev 2010;9:709-715.

22. National Committee for Clinical Laboratory Standards (NCCLS). Evaluation of Precision Performance of Quantitative Measurement Methods; Approved Guideline—Second Edition. NCCLS document EP5-A2 Wayne, PA: NCCLS, 2004.

23. National Committee for Clinical Laboratory Standards (NCCLS). Evaluation of the Linearity of Quantitative Measurement Procedures: A Statistical Approach; Approved Guideline. NCCLS document EP6-A. Wayne, PA: NCCLS; 2003.

24. National Committee for Clinical Laboratory Standards (NCCLS). Protocols for Determination of Limits of Detection and Limits of Quantitation; Approved Guideline. NCCLS document EP17-A, Wayne, PA: NCCLS, 2004.

25. Clinical and Laboratory Standards Institute. Interference Testing in Clinical Chemistry: Approved Guideline - Second Edition. CLSI Document EP7-A2. Wayne, PA: Clinical and Laboratory Standards Institute, 2005.

26. Passing HA, Bablok W. New biometrical procedure for testing the equality of measurements from two different analytical methods. J Clin Chem Clin Biochem 1983;21:709-20.

27. Wallace AM, Measurement of 25-hydroxyvitamin D in the clinical laboratory: Current procedures, performance characteristics and limitations. Steroids 2010;75:477-488.

The following US Patents are relevant to the ARCHITECT i System or its components. There are other such patents and patent applications in the United States and worldwide.

5 468 646 5 543 524 5 545 7395 565 570 5 669 819 5 783 699

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ARCHITECT and Chemiflex are trademarks of Abbott Laboratories in various jurisdictions.All trademarks are property of their respective owners.


Produced by Biokit S.A., 08186 Barcelona, Spain for Abbott

June 2011© 2010, 2011 Abbott Laboratories

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