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!