Author/Authors :
MOHAMMADIHA, Anita Dept. of Parasitology - Faculty of Medical Sciences - Tarbiat Modares University, Tehran, Iran , DALIMI, Abdolhossein Dept. of Parasitology - Faculty of Medical Sciences - Tarbiat Modares University, Tehran, Iran , MOHEBALI, Mehdi Center for Research of Endemic Parasites of Iran (CREPI) - Tehran University of Medical Sciences, Tehran, Iran , SHARIFI, Iraj Leishmaniasis Research Center - Dept. of Parasitology - Faculty of Medicine - Kerman University of Medical Sciences, Kerman, Iran , MAHMOUDI, Mohammadreza Dept. of Microbiology and Parasitology - Faculty of Medicine - Guilan University of Medical Sciences, Rasht, Iran , MIRZAEI, Asad Dept. of Parasitology - Faculty of Medicine - Ilam University of Medical Sciences, Ilam, Iran , SPOTIN, Adel Immunology Research Center - Tabriz University of Medical Sciences, Tabriz, Iran , BEHRAVAN, Mahmoodreza Esfarayen Faculty of Medical Sciences, Esfarayen, Iran , KARIMI, Mehdi Infectious Diseases Research Center - Microbiology Dept. - Birjand University of Medical Sciences, Birjand, Iran , ARBABI, Mohsen Dept. of Parasitology and Mycology, School of Medicine - Kashan University of Medical Sciences, Kashan, Iran , NEKOEIAN, Shahram Dept. of Cellular and Molecular Biology - School of Medicine - Isfahan University of Medical Sciences, Isfahan, Iran , KALANTARI, Reza Dept. of Parasitology - Faculty of Medical Sciences - Tarbiat Modares University, Tehran, Iran , GHORBANZADEH, Behzad Dept. of Parasitology - Faculty of Medical Sciences - Tarbiat Modares University, Tehran, Iran
Abstract :
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.
Keywords :
Phylogeny , L. major , L. tropica , PCR-RFLP , Human , Iran
