Title
An Innovative Field-Applicable Molecular Test to Diagnose Cutaneous Leishmania Viannia spp. Infections
Date Issued
26 April 2016
Access level
open access
Resource Type
journal article
Author(s)
Saldarriaga O.
Castellanos-Gonzalez A.
Porrozzi R.
Fernandez O.
Saravia N.
Costa E.
Melby P.
Travi B.
Unidad de Investigación Médica Naval-6
Unidad de Investigación Médica Naval-6
Publisher(s)
Public Library of Science
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.
Volume
10
Issue
4
Language
English
OCDE Knowledge area
Tecnologías que implican la manipulación de células, tejidos, órganos o todo el organismo
Dermatología, Enfermedades venéreas
Scopus EID
2-s2.0-84964798742
PubMed ID
Source
PLoS Neglected Tropical Diseases
ISSN of the container
19352727
Sponsor(s)
National Institute of Allergy and Infectious Diseases - R01AI093775.
Fogarty International Center - D43TW006589, D43TW007393.
Sources of information:
Directorio de Producción Científica
Scopus