بررسي پلي مورفيسم در ايزوله هاي گونه ليشمانيا ماژور تهيه شده در مناطق اندميك ليشمانيوز پوستي ايران با استفاده از ماركرهاي مايكروستلايت

Detection of Polymorphism within L. major isolates Collected from Iranian Patients Suffering from Cutaneous Leishmaniasis by Microsatellite Markers

Background and Objective: Protozoan parasites of Leishmania major is one of the causative agents of cutaneous leishmaniasis in different parts of Iran. Currently different molecular biology tools based on different polymorphic DNA sequences have been used in order to investigate heterogeneity among different Leishmania species. Multilocus microsatellite typing method (MLMT) is one of these interesting techniques that we applied to analyze leishmania major parasites collected from different endemic parts of zoonotic cutaneous Leishmaniasis (ZCL) in Iran. Materials and Methods: In this laboratory study, twenty-four leishmania major parasites collected from patients suffered from cutaneous leishmaniasis in different parts of Iran were investigated using MLMT method for the first time in Iran. The isolates were amplified by seven different designed primers for microsatellite markers and then analysed after running on polyacrylamide gel electrophoresis for detection of polymorphism among these isolates. Results: PCR products of 24 isolates using six microsatellite markers revealed polymorphic pattern, whereas monomorphic profile was observed with only one primer. Totally, seven different genotypes were demonstrated among studied isolates. Based on our results different genotypes were detected within leishmania major isolates collected from endemic areas of ZCL in Esfahan, Semnan and Tehran provinces, although only one genotype was observed in the studied isolated obtained from Khuzestan and Ilam foci. Conclusion: Overall, seven different genotypes were identified based on MLMT in studied Leishmania major isolates in different endemic areas of ZCL in Iran. Based on our finding We suggest that MLMT can utilised as a reliable method for detection of polymorphism within Leishmania major species.