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

Requena, Chicharro Carmen, García Lineth, Parrado Rudy, Puerta, Cañavate, Cañavate Carmen

Description

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 Hae III-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.

Sujets

• Leishmania
• Hsp70
• Three prime untranslated region
• Sequence analysis
• Microsatellite

Informations

• Publié par: biomed
• Publié le: 01 janvier 2012
• Nombre de lectures: 11
• Langue: English

Extrait

Requenaet al. Parasites & Vectors2012,5:87 http://www.parasitesandvectors.com/content/5/1/87

R E S E A R C H

Open Access

Sequence analysis of the 3’untranslated region of HSP70(type I) genes in the genusLeishmania: its usefulness as a molecular marker for species identification 1* 2 3 3 4 Jose M Requena , Carmen Chicharro , Lineth García , Rudy Parrado , Concepción J Puerta 2 and Carmen Cañavate

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 ofLeishmaniaspecies with excellent results. Methods:In the present study, we analyzed the 3’untranslated region (UTR) ofLeishmania HSP70type I gene from 24 strains representing elevenLeishmaniaspecies 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 subgenusLeishmaniaandViannia. In addition, the presence of many microsatellites was a common feature of the 3´UTR ofHSP70Igenes in theLeishmaniagenus. Finally, we constructed dendrograms based on global sequence alignments of the analyzedLeishmaniaspecies and strains, the results indicated that this particular region ofHSP70genes 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 theLeishmaniaandVianniasubgenera, direct visualization of the PCR amplification product would allow discrimination between subgenera, and aHaeIII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 WorldLeishmaniaspecies, might be useful as a molecular marker for species discrimination. Keywords:Leishmania, HSP70, 3’UTR, Sequence analysis, Microsatellites, Phylogenetic analysis

* Correspondence: [e-mail] 1 Centro de Biología Molecular“Severo Ochoa”(CSICUAM), Universidad Autonoma de Madrid, 28049 Madrid, Spain Full list of author information is available at the end of the article

© 2012 Requena et al.; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.