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Open Access Research

Sequence analysis of the 3’-untranslated region of HSP70 (type I) genes in the genus Leishmania: its usefulness as a molecular marker for species identification

Jose M Requena1*, Carmen Chicharro2, Lineth García3, Rudy Parrado3, Concepción J Puerta4 and Carmen Cañavate2

Author Affiliations

1 Centro de Biología Molecular “Severo Ochoa” (CSIC-UAM), Universidad Autonoma de Madrid, 28049, Madrid, Spain

2 WHO Collaborating Centre for Leishmaniasis, Servicio de Parasitología, Centro Nacional de Microbiología, Instituto de Salud Carlos III, 28220, Madrid, Spain

3 Instituto de Investigaciones Médicas, Fac. Medicina, Universidad Mayor de San Simón (UMSS), Cochabamba, Bolivia

4 Laboratorio de Parasitología Molecular, Facultad de Ciencias, Pontificia Universidad Javeriana, Bogotá, Colombia

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Parasites & Vectors 2012, 5:87  doi:10.1186/1756-3305-5-87

Published: 28 April 2012

Abstract

Background

The Leishmaniases are a group of clinically diverse diseases caused by parasites of the genus Leishmania. To distinguish between species is crucial for correct diagnosis and prognosis as well as for treatment decisions. Recently, sequencing of the HSP70 coding region has been applied in phylogenetic studies and for identifying of Leishmania species with excellent results.

Methods

In the present study, we analyzed the 3’-untranslated region (UTR) of Leishmania HSP70-type I gene from 24 strains representing eleven Leishmania species in the belief that this non-coding region would have a better discriminatory capacity for species typing than coding regions.

Results

It was observed that there was a remarkable degree of sequence conservation in this region, even between species of the subgenus Leishmania and Viannia. In addition, the presence of many microsatellites was a common feature of the 3´-UTR of HSP70-I genes in the Leishmania genus. Finally, we constructed dendrograms based on global sequence alignments of the analyzed Leishmania species and strains, the results indicated that this particular region of HSP70 genes might be useful for species (or species complex) typing, improving for particular species the discrimination capacity of phylogenetic trees based on HSP70 coding sequences. Given the large size variation of the analyzed region between the Leishmania and Viannia subgenera, direct visualization of the PCR amplification product would allow discrimination between subgenera, and a HaeIII-PCR-RFLP analysis might be used for differentiating some species within each subgenera.

Conclusions

Sequence and phylogenetic analyses indicated that this region, which is readily amplified using a single pair of primers from both Old and New World Leishmania species, might be useful as a molecular marker for species discrimination.

Keywords:
Leishmania; HSP70; 3’UTR; Sequence analysis; Microsatellites; Phylogenetic analysis