Please use this identifier to cite or link to this item: https://www.arca.fiocruz.br/handle/icict/7017
Title: Evaluation of 4 polymerase chain reaction protocols for cultured Leishmania spp. typing.
Authors: Rocha, Marcele Neves
Margonari, Carina
Presot, Ivanete Milagres
Soares, Rodrigo Pedro Pinto
Affilliation: Fundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Laboratory of Medical Entomology. Belo Horizonte, MG, Brazil.
Fundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Laboratory of Education and Health. Belo Horizonte, MG, Brazil.
Fundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Quality and Biosafety Section Belo Horizonte, MG, Brazil.
Fundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Laboratory of Medical Entomology. Belo Horizonte, MG, Brazil.
Abstract: Leishmaniasis is a disease caused by the protozoan Leishmania resulting in a variety of clinical manifestations, from self-healing skin lesions to fatal visceral disease. The development of polymerase chain reaction (PCR)-based techniques has made species identification easier, faster, and less labor intensive. The main targets for PCR amplification include kinetoplastid DNA (kDNA), miniexon, and conserved regions such as the internal transcribed spacer. The objective of this work was to evaluate 4 different PCR techniques designed to type Leishmania using laboratory strains. Parasites were subjected to 4 PCR procedures using specific Leishmania primers for miniexon (designated A1 and A2) and kDNA (designated B1 and B2, C1 and C2, and D1, D2 and D3). Discrimination between some species and the 2 main subgenera Leishmania and Viannia was achieved. Unweighted pair group method analysis resulted in the expected clustering of the 2 species from the subgenus Leishmania. However, some species in the subgenus Viannia could not be distinguished, representing a continued challenge for PCR-based protocols. Results are discussed in terms of advantages, limitations, and reproducibility of these 4 PCR-based techniques in the taxonomy of Leishmania.
Keywords: Polymerase chain reaction
Leishmania
Typing
Diagnosis
Issue Date: 2010
Publisher: Elsevier Biomedica
Citation: ROCHA, Marcele Neves et al. Evaluation of 4 polymerase chain reaction protocols for cultured Leishmania spp. typing. Diagn Microbiol Infect Dis., v. 68, n. 4, p. 401-409, Dec. 2010.
DOI: 10.1016/j.diagmicrobio.2010.08.007
Copyright: restricted access
Appears in Collections:MG - IRR - Artigos de Periódicos

Files in This Item:
File Description SizeFormat 
96.2010.pdf586.46 kBAdobe PDFThumbnail
    Request a copy



FacebookTwitterDeliciousLinkedInGoogle BookmarksBibTex Format mendeley Endnote DiggMySpace

Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.