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Background and purpose: Leishmaniasis is an important tropical disease in Iran and the world. Despite extensive effort to make a vaccine, no success was achieved. Moreover, routine methods have failed to control and cut the spread of this disease. Hence, the major aim of this study was to identify and determine the Leishmania infections in the vector and the ecological and biological criteria of the vector by using new molecular methods to regionally control leishmaniasis. Materials and methods: Head and abdominal terminalia of the sandflies collected on sticky papers and CDC traps were dissected and mounted on the slide. Then, the species were determined using microscope and morphological keys. Furthermore, DNA was extracted from the dissected thorax and attached anterior abdomen of the sandflies. Leishmania parasite was detected and identified using digestion of restriction enzyme (RFLP) and sequencing of ITS-rDNA gene. Results: Out of 168 Phlebotomus papatasi, 18 had Leishmania infection. Two species of Leishmania major and Leishmania turanica were identified in Phlebotomus papatasi in Turkmen Sahara. Identification of these two parasites was confirmed after amplifying ITS-rDNA gene using Nested PCR, RFLP and sequencing. Conclusion: Using new molecular methods, it was reconfirmed that Leishmania major was the causative agent and Phlebotomus papatasi was a vector of rural zoonotic cutaneous leishmaniasis (ZCL) in Turkmen Sahara and Iran. Finding Leishmania turanica in Phlebotomus papatasi and in reservoirs can indicate the role of this parasite in the durability and stability of the disease cycle.

Keywords: Leishmania major, Leishmania turanica, Phlebotomus papatasi, ITS-ribosomal DNA

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