Molecular identification of Leishmania species isolated from patients with cutaneous leishmaniosis in gonbad Kavoos, northeastern of Iran using hSP70 and ITS-based PCR-RFlP

Cutaneous leishmaniosis (CL) is mainly caused by Leishmania major (rural-type) and Leishmania tropica (urban-type). CL is a major health problem in many regions of the world, and it is associated with health complications and economic loss. The identification and differentiation of Leishmania species are critical because the prevention and control methods, as well as management and therapeutic strategies, are different for each type of CL. The present study aimed to identify the parasite species responsible for CL in the study area using ITS1 and HSP70- based PCR-RFLP methods. A total of 147 stained slides were prepared from samples collected from CL patients, and these slides were positive for amastigotes of Leishmania species on microscopic examination. Forty-three Giemsastained slides with 2+ to 4+ grades were selected for molecular studies for the identification of the Leishmania species. DNA was extracted from the selected slides for the molecular studies. The amplification of HSP70 and ITS1 genes was performed by the PCR method. The PCR products were digested with the HaeIII restriction enzyme, and banding patterns of all samples were compared with reference strains. Overall, patterns of all the samples were found to correspond to the reference strains of L. major based on RFLP-PCR targeting HSP70 and ITS1 genes of the parasite, demonstrating the dominance of L. major as the causative agent of zoonotic cutaneous leishmaniosis (zCL) in the study area. This area is endemic for zoonotic CL, and further studies are required to determine the reservoir and natural infection of sand flies in this county.