An Innovative Field-Applicable Molecular Test to Diagnose Cutaneous Leishmania Viannia spp. Infections

Omar A. Saldarriaga, Alejandro Castellanos, Renato Porrozzi, Gerald C. Baldeviano, Andrés G. Lescano, Maxy B. de Los Santos, Olga L. Fernandez, Nancy G. Saravia, Erika Costa, Peter Melby, Bruno Travi

Research output: Contribution to journalArticle

13 Scopus citations

Abstract

Cutaneous and mucosal leishmaniasis is widely distributed in Central and South America. Leishmania of the Viannia subgenus are the most frequent species infecting humans. L. (V.) braziliensis, L. (V.) panamensis are also responsible for metastatic mucosal leishmaniasis. Conventional or real time PCR is a more sensitive diagnostic test than microscopy, but the cost and requirement for infrastructure and trained personnel makes it impractical in most endemic regions. Primary health systems need a sensitive and specific point of care (POC) diagnostic tool. We developed a novel POC molecular diagnostic test for cutaneous leishmaniasis caused by Leishmania (Viannia) spp. Parasite DNA was amplified using isothermal Recombinase Polymerase Amplification (RPA) with primers and probes that targeted the kinetoplast DNA. The amplification product was detected by naked eye with a lateral flow (LF) immunochromatographic strip. The RPA-LF had an analytical sensitivity equivalent to 0.1 parasites per reaction. The test amplified the principal L. Viannia species from multiple countries: L. (V.) braziliensis (n = 33), L. (V.) guyanensis (n = 17), L. (V.) panamensis (n = 9). The less common L. (V.) lainsoni, L. (V.) shawi, and L. (V.) naiffi were also amplified. No amplification was observed in parasites of the L. (Leishmania) subgenus. In a small number of clinical samples (n = 13) we found 100% agreement between PCR and RPA-LF. The high analytical sensitivity and clinical validation indicate the test could improve the efficiency of diagnosis, especially in chronic lesions with submicroscopic parasite burdens. Field implementation of the RPA-LF test could contribute to management and control of cutaneous and mucosal leishmaniasis.

Original languageEnglish (US)
Article numbere0004638
JournalPLoS Neglected Tropical Diseases
Volume10
Issue number4
DOIs
StatePublished - Apr 26 2016

    Fingerprint

ASJC Scopus subject areas

  • Infectious Diseases
  • Public Health, Environmental and Occupational Health
  • Pharmacology, Toxicology and Pharmaceutics(all)

Cite this

Saldarriaga, O. A., Castellanos, A., Porrozzi, R., Baldeviano, G. C., Lescano, A. G., de Los Santos, M. B., Fernandez, O. L., Saravia, N. G., Costa, E., Melby, P., & Travi, B. (2016). An Innovative Field-Applicable Molecular Test to Diagnose Cutaneous Leishmania Viannia spp. Infections. PLoS Neglected Tropical Diseases, 10(4), [e0004638]. https://doi.org/10.1371/journal.pntd.0004638