Real-time PCR to quantify Leishmania donovani in hamsters

Anuradha Srivastava, J. Mark Sweat, Azliyati Azizan, Brian Vesely, Dennis E. Kyle

Research output: Contribution to journalArticlepeer-review

13 Citations (Scopus)

Abstract

Visceral leishmaniasis, a vector-borne disease caused by Leishmania donovani and Leishmania infantum, currently affects 12 million individuals in 88 countries. In the present study, a real-time PCR (rt-PCR) assay has been optimized and validated against 2 other routine methods, i.e., microscopy and limiting dilution culture assay, to estimate parasite load in the liver of infected Syrian hamsters (Mesocricetus auratus). A set of specific primers amplified a 116-bp target template of the kinetoplastid DNA of L. donovani in a SYBR® Green-based rt-PCR assay. To assess the methods, we tested 2 anti-leishmanial compounds belonging to the class of arylimidamides, DB745 (2,5-bis[2-ethoxy-4-(2-pyridylimino)aminophenyl]furan) and DB766 (2,5-bis[2-(2-propoxy)-4-(2-pyridylimino)aminophenyl]furan) for efficacy in vivo in Syrian hamsters infected with L. donovani promastigotes. Parasite load was quantified in liver by all 3 methods and was found comparable. Of the 3 methods, rt-PCR was the fastest and most convenient, sensitive, and reproducible method.

Original languageEnglish
Pages (from-to)145-150
Number of pages6
JournalJournal of Parasitology
Volume99
Issue number1
DOIs
Publication statusPublished - Feb 2013

ASJC Scopus subject areas

  • Parasitology
  • Ecology, Evolution, Behavior and Systematics

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