Validation of a simple resazurin-based promastigote assay for the routine monitoring of miltefosine susceptibility in clinical isolates of Leishmania donovani

A. Kulshrestha, V. Bhandari, R. Mukhopadhyay, V. Ramesh, S. Sundar, L. Maes, J.C. Dujardin, S. Roy, P. Salotra

Research output: Contribution to journalA1: Web of Science-article

Abstract

Simple, cost-effective approach for routine surveillance of parasite susceptibility to antileishmanial drug miltefosine (MIL) is highly desirable for controlling emergence of drug resistance in visceral leishmaniasis (VL). We validated a simple resazurin-based fluorimetric assay using promastigotes to track natural MIL tolerance in Leishmania donovani parasites from VL cases (n = 17) against standard amastigote assay, in two different labs in India. The inter-stage MIL susceptibility correlated strongly (r = 0.70, p = 0.0018) using J774.A.1 macrophage cell line-based amastigote assay and fluorescence-based resazurin assay for promastigotes. Investigation of inter-stage MIL susceptibility for the same set of clinical isolates in another lab also showed a strong correlation (r = 0.72, p = 0.0012) using mouse peritoneal macrophages for amastigote assay and resazurin-based alamar blue assay for promastigotes. Additionally, parasites from post-kala-azar dermal leishmaniasis (PKDL) lesions (n = 7, r = 0.78, p = 0.046) and MIL-induced parasites (r = 0.92, p = 0.0001; n = 3) also exhibited a strongly correlated inter-stage miltefosine susceptibility. Thus, our results support the utility of resazurin assay as a simplified biological tool for MIL susceptibility monitoring in clinical isolates from MIL-treated VL/PKDL patients.
Original languageEnglish
JournalParasitology Research
Volume112
Issue number2
Pages (from-to)825-828
Number of pages4
ISSN0932-0113
DOIs
Publication statusPublished - 2013

Keywords

  • Protozoal diseases
  • Kala azar
  • Visceral
  • Leishmaniasis
  • Leishmania donovani
  • Vectors
  • Sandflies
  • Phlebotomus argentipes
  • Drug susceptibility
  • Miltefosine
  • Routine
  • Monitoring
  • Surveillance
  • Validation
  • Resazurin
  • Assays
  • Promastigotes
  • Laboratory techniques and procedures
  • India
  • Asia-South

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