Characterization of leishmania species and L. major stains in different endemic areas of cutaneous leishmanasis Iran

ABSTRACT

Both zoonotic and anthroponotic cutaneous leishmaniasis [CL] caused by L. major and L. tropica, respectively, are endemic in different parts of Iran. This study was performed to investigate the new changes in epidemiological pattern of CL, and to identify the species of Leishmania and the strains of L. major isolated from different endemic areas of Iran. Seventy-two isolates from 245 samples collected from different endemic areas of Iran were characterized by monoclonal antibodies [mAb] specific for L. major, L. tropica, and L. infantum. Flow cytometry, isoenzyme analysis and PCR amplification were used for identification. Isoenzyme analysis was carried out by PAGE and cellulose acetate. Eight enzymes including malate dehydrogenase [MDH], malic enzyme [ME], glucose 6-phosphate dehydrogenase [G6PD], glucose phosphate isomerase [GPI], nucleoside hydrolase inosine [NHi], nucleoside hydrolase deoxy inosine [NHd], superoxide dismutase [SOD] and 6-phosphogluconate dehydrogenase [6PGD] were used for isoenzyme analysis. PCR assay was developed using specific primers against kinetoplast DNA. The isolates were compared with reference strains [RS]. Data obtained from different methods showed 45 out of 72 isolates were similar to RS of L. major and 22 similar to L. tropica, and also five samples were identified as Crithidia. Isoenzyme migration pattern of L. major isolates was indistinguishable from each other in six enzymatic systems but differences were observed in profile of two enzymes of SOD and MDH. The data indicate that L. major species are dominant in the studied endemic areas, and different strains of L. major are present in different geographic areas of Iran. Moreover, it seems that enzymatic system is more useful approach than others for characterization of Leishmania species and L. major strains