Molecular Characterization of Leishmania Infection in Sand flies From Sistan Va Baluchistan Province, Southeastern Iran

Background: Cutaneous leishmaniasis (CL) is a zoonotic disease that is caused by various species of the genus Leishmania. The disease is considered a major health problem in different areas of Iran and is an endemic disease in rural areas of Mirjaveh, Chabahar, and Konarak Counties, Sistan Va Baluchistan Province. Objectives: The aim of this study was to identify Leishmania species that was isolated from potential sand fly vectors by molecular analysis in Chabahar County. Materials and Methods: To collect Sand flies, sticky traps were placed at the entrance of rodents burrows in Dashtiyari division of Chabahar County, where CL is endemic. Freshly collected Sand flies were identified with regard to species, dissected in normal saline using binocular, and examined for leptomonads under a microscope. Leptomonads from the Sand flies were used to inoculate the base of Balb/c mice tails subcutaneously; after an incubation period and the development of lesions, the parasites were transferred to NNN + LIT medium culture. The harvested Leishmania parasites were subjected to DNA extraction and analyzed by random amplified polymorphic DNA polymerase chain reaction (RAPD-PCR). Results: DNA from Leishmania species from Phlebotomus papatasi and P. salehi Sand flies produced distinctive patterns of bands of L. major with all primers. However, the products at approximately 2100 bp and 800 bp that were amplified with primer 329 were stable and re- producible in all assays. This is the first report on the isolation and identification of L. major in P. salehi from Iran and P. papatasi from Sistan va Baluchistan. Conclusions: The study shows that P. papatasi and P. salehi Sand flies play a major role in the maintenance and transmission of disease to humans in this area.