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Open Access Research article

Polymorphism-specific PCR enhances the diagnostic performance of American tegumentary leishmaniasis and allows the rapid identification of Leishmania species from Argentina

Jorge D Marco12*, Paola A Barroso12, Tatsuyuki Mimori3, Fabricio M Locatelli2, Ayako Tomatani4, María C Mora1, S Pamela Cajal5, Julio R Nasser5, Luis A Parada1, Taketoshi Taniguchi4, Masataka Korenaga2, Miguel A Basombrío1 and Yoshihisa Hashiguchi26

Author Affiliations

1 Instituto de Patología Experimental, Facultad de Ciencias de la Salud, Universidad Nacional de Salta / Consejo Nacional de Investigaciones Científicas y Técnicas, Salta, Argentina

2 Department of Parasitology, Kochi Medical School, Kochi University, Nankoku, Japan

3 Department of Microbiology, School of Health Sciences, Kumamoto University, Kumamoto, Japan

4 Division of Molecular Biology, Science Research Center, Kochi University, Nankoku, Japan

5 Instituto de Investigaciones en Enfermedades Tropicales, Sede Regional Orán, Universidad Nacional de Salta, San Ramón de la Nueva Orán, Salta, Argentina

6 Prometeo Proyecto, Centro de Biomedicina, Universidad Central del Ecuador, Quito, Ecuador

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BMC Infectious Diseases 2012, 12:191  doi:10.1186/1471-2334-12-191

Published: 15 August 2012

Abstract

Background

The diagnosis of the leishmaniases poses enormous challenges in Argentina. The Polymorphism-Specific PCR (PS-PCR) designed and validated in our laboratories has been proven effective for typifying the Leishmania genus from cultured material. Here we evaluated the performance of this method in the diagnosis of American tegumentary leishmaniasis (ATL) and the rapid identification of Leishmania spp. directly from clinical specimens.

Methods

A total of 63 patients from northwestern Argentina, with cutaneous or mucocutaneous lesions, underwent an ATL diagnosis protocol which included clinical examination, Leishmanin skin test, and microscopic examination of dermal smears. In addition, we performed PS-PCR on DNA directly extracted from the specimens scraped from the lesions.

Results

Out of the 63 patients, 44 were classified as ATL cases and 19 as non-ATL cases. The diagnostic sensitivity of the microscopic analysis of dermal smears and PS-PCR individually were 70.5% and 81%, respectively. When performing both tests in parallel, this parameter increased significantly to 97.6% (p = 0.0018). The specificities, on the other hand, were 100%, 84.2%, and 83.3% for the combination, respectively (p > 0.05). Using the PS-PCR analysis we successfully identified the Leishmania spp. in 31 out of the 44 ATL cases. Twenty-eight (90.3%) cases were caused by L. (V.) braziliensis, two (6.5%) by L. (V.) guyanensis, and one (3.2%) by L. (V.) panamensis.

Conclusions

The efficacy of the ATL diagnosis was significantly improved by combining the dermal smear examination with a PS-PCR analysis. Our strategy allowed us to reach the diagnosis of ATL with high accuracy regarding the species of the etiological agent in 70.5% of the cases. Moreover, we diagnosed two cases of the disseminated cutaneous form caused by L. (V.) braziliensis and a cutaneous case due to L. (V.) panamensis infection, both findings reported for the first time in Argentina.

Keywords:
Leishmaniasis; Tegumentary leishmaniasis; Leishmania braziliensis; Leishmania guyanensis; Leishmania panamensis; PS-PCR; Diagnosis; Argentina; Species identification; Leishmania strains