n Beheshti - Parasitology Department, Medical Sciences Faculty, Tarbiat Modares University, Tehran, Iran
f Ghaffarifar - Parasitology Department, Medical Sciences Faculty, Tarbiat Modares University, Tehran, Iran
z Sharifi - Blood Transfusion Research Center, High Institute for Research & Education in Transfusion Medicine, Tehran, Iran
z Eslamirad - Parasitology Department, Medical Sciences Faculty, Arak University of Medical Sciences, Arak, Iran
m Farivar Sadri - Parasitology Department, Medical Sciences Faculty, Tehran University of Medical Sciences, Tehran, Iran
m.s Dayer - Parasitology Department, Medical Sciences Faculty, Tarbiat Modares University, Tehran, Iran

Aims The aim of this study was species identification and phylogenic analysis of species of Leishmania isolated from clinical samples. Materials & Methods The samples were collected from patients that were infected from different parts of Iran. After microscopic examination, we used PCR method for amplify the ITS1 (internal transcribed spacer 1) gene. RFLP method (digestion with Apo1 restriction enzyme) and for phylogenetic construction, DNA sequencing of PCR product were used. Findings Two samples from Khorasan province (Mashhad) were Leishmania tropica (L. tropica), while others were Leishmania major (L. major). L. tropica samples are more variable compared with L. major. The molecular sequencing differences between L. major was related to geographical distribution. Based on the results of PCR product in the gel electrophoresis and DNA sequencing for L. tropica and L. major, the DNA sizes were between 350 and 369bp. The RFLP for L. major and L. tropica showed two and one bands, respectively. The sequences for all samples from central parts are the same, but there is difference with the samples isolated from North-East part of Iran. Conclusion The sequences of ITS1 gene of Leishmania major separated from Damghan and Esfarayen are different from other samples. Similarity of DNA sequences of North-East part of Iran of L. major with samples from central parts was 99%. The similarity of two isolates of L. tropica was 96%. The most similarity of Leishmania tropica isolated was 95% with Indian isolate and the most similarity for Leishmania major was 99% with Friedlin strain

Leishmania Major; PCR; RFLP; Sequencing; Phylogenetic Analysis; Iran