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

MARCO, JORGE DIEGO; UEZATO, HIROSHI; MIMORI, TATSUYUKI; BARROSO, PAOLA ANDREA; KORENAGA, MASATAKA; NONAKA, SHIGEO; BASOMBRÍO, MIGUEL ANGEL; TARANTO, NÉSTOR JUAN; HASHIGUCHI, YOSHIHISA

AMERICAN JOURNAL OF TROPICAL MEDICINE AND HYGIENE

American Society of Tropical Medicine and Hygiene

Lugar: Cleveland, United States of America; Año: 2006 vol. 75 p. 256 - 260

0002-9637

Abstract. 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 wereassigned 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 thecountry