A novel molecular test to diagnose canine visceral leishmaniasis at the point of care

Alejandro Castellanos, Omar A. Saldarriaga, Lilian Tartaglino, Rosana Gacek, Elissa Temple, Hayley Sparks, Peter Melby, Bruno Travi

Research output: Contribution to journalArticle

17 Scopus citations

Abstract

Dogs are the principal reservoir hosts of zoonotic visceral leishmaniasis (VL) but current serological methods are not sensitive enough to detect all subclinically infected animals, which is crucial to VL control programs. Polymerase chain reaction (PCR) methods have greater sensitivity but require expensive equipment and trained personnel, impairing its implementation in endemic areas. We developed a diagnostic test that uses isothermal recombinase polymerase amplification (RPA) to detect Leishmania infantum. This method was coupled with lateral flow (LF) reading with the naked eye to be adapted as a point-of-care test. The L. infantum RPA-LF had an analytical sensitivity similar to real time-PCR, detecting DNA of 0.1 parasites spiked in dog blood, which was equivalent to 40 parasites/mL. There was no cross amplification with dog or human DNA or with Leishmania braziliensis, Leishmania amazonensis, or Trypanosoma cruzi. The test also amplified Leishmania donovani strains (N = 7). In a group of clinically normal dogs (N = 30), RPA-LF detected more subclinical infections than rK39 strip test, a standard serological method (50% versus 13.3% positivity, respectively; P = 0.005). Also, RPA-LF detected L. infantum in noninvasive mucosal samples of dogs with a sensitivity comparable to blood samples. This novel molecular test may have a positive impact in leishmaniasis control programs.

Original languageEnglish (US)
Pages (from-to)970-975
Number of pages6
JournalAmerican Journal of Tropical Medicine and Hygiene
Volume93
Issue number5
DOIs
StatePublished - Nov 4 2015

ASJC Scopus subject areas

  • Parasitology
  • Infectious Diseases
  • Virology

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