Are cytochrome B gene sequencing and polymorphism-specific polymerase chain reaction as reliable as multilocus enzyme electrophoresis for identifying Leishmania spp. from Argentina?

Recently, two techniques, polymerase chain reaction (PCR) amplification and sequencing of cytochrome b gene (cyt b gene sequencing) and polymorphism-specific PCR (PS-PCR) were recommended for Leishmania species identification. Before this study, however, the accuracy of these methods had not been te...

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Bibliographic Details
Published in:	The American journal of tropical medicine and hygiene Vol. 75; no. 2; p. 256
Main Authors:	Marco, Jorge D, Uezato, Hiroshi, Mimori, Tatsuyuki, Barroso, Paola A, Korenaga, Masataka, Nonaka, Shigeo, Basombrío, Miguel A, Taranto, Néstor J, Hashiguchi, Yoshihisa
Format:	Journal Article
Language:	English
Published:	United States 01-08-2006
Subjects:	Animals Argentina Base Sequence Cytochromes b - genetics DNA Primers - chemistry Double-Blind Method Electrophoresis, Agar Gel Leishmania - classification Leishmania - enzymology Leishmania - genetics Molecular Sequence Data Polymerase Chain Reaction - methods Polymerase Chain Reaction - standards Polymorphism, Genetic Reproducibility of Results Sequence Alignment - methods Species Specificity Argentina
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Summary:	Recently, two techniques, polymerase chain reaction (PCR) amplification and sequencing of cytochrome b gene (cyt b gene sequencing) and polymorphism-specific PCR (PS-PCR) were recommended for Leishmania species identification. Before this study, however, the accuracy of these methods had not been tested against the multilocus enzyme electrophoresis, the current gold standard technique on this task. Therefore, a trial was done for the first time to compare the results obtained by these techniques, using 17 Argentinean Leishmania stocks in independent assays. For all the stocks examined, the same results at species level were obtained by the three techniques. Among them, 14 were assigned to L. (Viannia) braziliensis, and three to L. (V.) guyanensis. The two techniques, cyt b gene sequencing and PS-PCR, were able to distinguish between all the proven species responsible for leishmaniases in Argentina. Thus, both techniques were validated and could be used independently for the species designation of Leishmania parasites in the country.
ISSN:	0002-9637
DOI:	10.4269/ajtmh.2006.75.256