Author/Authors :
Beheshti, N. Parasitology Department - Medical Sciences Faculty - Tarbiat Modares University, Tehran, Iran , Ghaffari Far, F. Parasitology Department - Medical Sciences Faculty - Tarbiat Modares University, Tehran, Iran , Sharifi, Z. Blood Transfusion Research Center - High Institute for Research & Education in Transfusion Medicine, Tehran, Iran , Eslamirad, Z. Parasitology Department - Medical Sciences Faculty - Arak University of Medical Sciences, Arak, Iran , Farivar Sadr, M. Parasitology Department - Medical Sciences Faculty - Tehran University of Medical Sciences, Tehran, Iran , Dayer, M.S. Parasitology Department - Medical Sciences Faculty - Tarbiat Modares University, Tehran, Iran , Nasiri, V. Parasitology Department - Razi Vaccine & Serum Research Institute - Agricultural Research, Education and Extension Organization (AREEO), Karaj, Alborz, Iran , Ebrahimisadr, P. Parasitology Department - Medical Sciences Faculty - Tarbiat Modares University, Tehran, Iran , Hamidianfar, N. Natural Resources & Environmental Sciences Department - Agriculture & Natural Resources Faculty, Khorasgan (Isfahan) Branch, Islamic Azad University Isfahan, Iran , Bayesh, S. Students Research Committee - Tehran Medical Sciences Branch, Islamic Azad Univesity, Tehran, Iran , Jorjani, O Laboratory Science Research Center - Golestan University of Medical Sciences, Gorgan, Iran
Abstract :
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
Keywords :
Iran , Phylogenetic Analysis , Sequencing , RFLP , Leishmania Major , PCR
