Parasitology. Accuracy of cytochrome b gene sequencing and polymorphism specific polymerase chain reaction on the species discrimination of Argentinean Leishmania spp., characterized by multilocus enzyme electrophoresis

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

Studies on New and Old World Leishmaniasis and their transmission, with particular reference to Ecuador, Argentina and Pakistan

Kyowa Printing & Co. Ltd.

Lugar: Kochi, Japan; Año: 2004; p. 22 - 28

The Leishmania species identification is important clinico-epidemiologically in all the areas endemic for the diseases. Recently, DNA based techniques have been developed for this purpose. In the present study, the species discrimination of 17 Argentinean stocks isolated from human cutaneous, mucocutaneous, recurrent and canine cutaneous leishmaniasis cases, previously characterized by multilocus enzyme electrophoresis (MLEE), was done by Cytochrome b gene sequencing (Cyt b) and polymorphism-specific polymerase chain reaction (PS-PCR) techniques in a triple blind assay. For all the stocks, the same results were obtained by Cyt b and MLEE.  These techniques were therefore validated. Fourteen of 17 stocks were assigned to Leishmania (Viannia) braziliensis, and the remaining three, to L. (V.) guyanensis by the two techniques Cyt b and MLEE. Although the PS-PCR protocol was also validated  in the discrimination of the 14 L. (V.) braziliensis stocks, it determined the rest as L. (V.) panamensis including all the L. (V.) guyanensis stocks assigned by Cyt b and MLEE. Thus, these results must be interpreted as L. (V.) guyanensis/panamensis, since the PS-PCR protocol could not distinguish between them at species level. The two techniques, Cyt b and PS-PCR, were able to distinguish all the proven species responsible for American tegumentary leishmaniasis in the northern areas of Argentina. Besides, for PS-PCR protocol it is not necessary to isolate the parasites for its application using cotton swabs and others. Therefore, PS-PCR would be useful for the analysis of a large number of samples from the disease endemic areas.