Abstract

CHENA, L et al. Characterization of Leishmania strains using PCR-RFLP of Spliced Leader Miniexon(SLME) in human and canine isolates in Paraguay. Mem. Inst. Investig. Cienc. Salud [online]. 2012, vol.10, n.1, pp.14-23. ISSN 1812-9528.

Leishmaniasis is a parasitic disease with various clinical forms from minor skin lesions to fatal diseases with visceral compromise. There are several molecular techniques such as the polymerase chain reaction (PCR) that amplifies specific target sequences like the SLME(spliced leader miniexon) region. Subsequently, the PCR products are digested with specific restriction enzymes (PCR-RFLP) allowing the identification of Leishmania species. This study was carried out in order to characterize Leishmania strains by using a PCR-RFLP of the SLME region in human and canine isolates from different regions of the country. We analyzed 12 human isolates and 40 dog isolates properly codified. Two primers for SLME region were used and the amplified products were digested with the Hae III and Nco I (RFLP) enzymes, then the banding patterns were analyzed. In 7 human isolates, parasites from the Viannia subgenus were detected while in 5 human isolates and 40 dog isolates, parasites from the Leishmania subgenus were identified. RFLP, according to the restriction banding patterns, allowed the identification of L. braziliensis species in the tegumentary leishmaniasis isolates and L. chagasi in visceral leishmaniasis isolates. This is the first study conducting molecular characterization of Leishmania species in the country, and its findings have implications at the epidemiological level by contributing to the improvement of surveillance strategies for the disease control. Additionally, the use of other genetic markers is suggested in order to identify genotypes or different genetic profiles among the strains of these species.

Keywords : Leishmania; genotyping; PCR-RFLP; miniexon.

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