Tehran University of Medical Sciences Iranian Journal of Parasitology 1735-7020 13 3 2018 09 23 351 361 EN Anita MOHAMMADIHA Dept. of Parasitology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran Abdolhossein DALIMI Dept. of Parasitology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran Mehdi MOHEBALI Dept. of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran AND Center for Research of Endemic Parasites of Iran (CREPI), Tehran University of Medical Sciences, Tehran, Iran Iraj SHARIFI Leishmaniasis Research Center, Dept. of Parasitology, Faculty of Medicine, Kerman University of Medical Sciences, Kerman, Iran Mohammadreza MAHMOUDI Dept. of Microbiology and Parasitology, Faculty of Medicine, Guilan University of Medical Sciences, Rasht, Iran Asad MIRZAEI Dept. of Parasitology, Faculty of Medicine, Ilam University of Medical Sciences, Ilam, Iran Adel SPOTIN Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran Mahmoodreza BEHRAVAN Esfarayen Faculty of Medical Sciences, Esfarayen, Iran Mehdi KARIMI Infectious Diseases Research Center, Microbiology Dept., Birjand University of Medical Sciences, Birjand, Iran Mohsen ARBABI Dept. of Parasitology and Mycology, School of Medicine, Kashan University of Medical Sciences, Kashan, Iran Shahram NEKOEIAN Dept. of Cellular and Molecular Biology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran Reza KALANTARI Dept. of Parasitology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran Behzad GHORBANZADEH Dept. of Parasitology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran 2017 02 08 2017 07 18 Background: In Iran, both forms of cutaneous (CL) and visceral leishmaniasis (VL) have been reported; so the accurate species identification of the parasite(s) and the analysis of genetic diversity are necessary. Methods: The smears were collected from lesions samples of 654 patients with CL, who attended local health centers in 12 provinces of Iran during 2013-2015. The smears were checked for the presence of amastigotes by light microscopy. DNA of 648 Leishmania isolates, amplified by targeting a partial sequence of ITS (18S rRNA–ITS1–5.8S rRNA–ITS2) gene. Twenty-five of all the amplicons were sequenced and analyzed with restriction fragment length polymorphism (RFLP) using the Taq1 enzyme. Results: All the smears were positive microscopically. The PCR-RFLP analysis revealed that 176 (27%) CL patients were infected with L. tropica and, 478 (73%) with L. major. The dominant species in all over Iran is L. major. The sequencing results of all CL patients and RFLP analysis confirmed each other. Based on our phylogenetic tree, 25 ITS DNA sequences were grouped into two clusters representing L. major and L. tropica species. Phylogenetic tree derived from the ITS sequences supports a clear divergence between L. major from the other species. Conclusion: Discrimination of Iranian Leishmania isolates using ITS gene gives us this opportunity to detect, identify, and construct the phylogenetic relationship of Iranian isolates. https://ijpa.tums.ac.ir/index.php/ijpa/article/view/1479 https://ijpa.tums.ac.ir/index.php/ijpa/article/download/1479/848