leishmaniasis biología molecular
TRANSCRIPT
Presented by: - DAVID GAVIRIA ORTIZ -ANA MARÍA GÓMEZ GÓMEZ
BODY OF OUR PRESENTATION INTRODUCTION.AIM OF THE STUDY.METHODS.RESULTS.DISCUSSION.CLOSURES.CONCEPT MAPS.
INTRODUCTION
GENETIC MARKERS
LEISHMANIASIS
DRUG RESISTAN
CE
• Female phlebotomine sandflies
• Promastigote
• Amastigote
L I F E C Y C L E
GENERAL OBJECTIVECharacterize each clinical
isolate of Indian KA phenotypes
genotypes
attempting to the development of markers for drug resistance,
if any.
MATERIALES Y MÉTODOS
DECLARACIONES ÉTICAS
CULTIVO DE PARÁSITOS DE LEISHMANIA Y CEPAS DE
REFERENCIA
Cepas de referencia de- L. donovani (DD8)- L. tropica (K27)- L. donovani
(MHOM/IN/1983/AG83)
CEPAS CONTROL
Amastigotes confirmados con Giemsa.
Haga clic en el icono para agregar una imagen
https://www.cdc.gov/dpdx/leishmaniasis/
AISLAMIENTO DE DNA GENÓMICO
PCR-RFLP
• Polimorfismos.•Enzimas de restricción.
HaeIII
MURINE MACROPHAGE LIKE TUMOR
CELL
ENSAYO DE SUSCEPTIBILIDAD
• Para ver si hay resistencia o sensibilidad a una droga.
L. donovani
SSG
MacrófagosPromastigo
tes
SSCP
• Detectar mutaciones y genotipar.• 4 pasos.
Fragmentos de control
• L0110• L510• L413• LK413• L0308• L0114
Fragmentos usados
• ITS1• ITS2
RESULTADOS
SSG Estibogluconato de sodio
SSCPPolimorfismo conformacional de hebra sencilla
ITS1
hsp70
S
S
R
R
AI: >3.0 R <3.0 S
ITS1 A B C D
L. donovani
L. tropicaSSG-R
ITS2 X Y Z
L. donovani
L. tropica SSG-R
Fragmentos anónimos de DNA
PCR
especies
SSG R – S
QRS
≠ Patrones polimórficosSSG R
DISCUSSION
AUTHOR WHAT THEY SAID OUTCOMESSCP is one of the basic methods to identify single base divergence in genes.
Gasser
Understanding the molecular mechanisms of clinical resistance is crucial for analyse the progress and extent of drug resistance in the field.
Asutosh
Srivastava Significant genetic polymorphisms exist in L. donovani strains in Bihar, India.You can differenciate resistant and sensitive SSG Leishmania parasites, by protein encoding genes, rDNA and mini-exon.
Downing
CLOSURES• For this pathology, it isn’t enough for the sandflies to
transmit the parasites just by a bite, it is obligatory an intra-macrophage process.
• One can tell apart the resistant Leishmania isolates from the sensitive Leishmania isolates on the SSCP method.
• In order to avoid drug resistance, we need to understand the polymorphisms regarding each parasite.
• Of the few samples that were positive for the codification of L. tropica, they could recognize the susceptibility for the SSG. Discovering an important, but not of huge magnitude diagnosis impression.
ANA’S CONCEPT MAP
DAVID´S CONCEPT MAP
Thank you!