Molecular Identification and Phylogenetic Classification of Leishmania spp. Isolated from Human Cutaneous Leishmani-asis in Iran: A Cross-sectional Study

  • 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 3. 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
Keywords: Phylogeny, L. major, L. tropica, PCR-RFLP, Human, 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.

Author Biographies

Anita MOHAMMADIHA, Dept. of Parasitology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran
Department of Parasitology, Medical Sciences Faculty
Mehdi MOHEBALI, Dept. of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran 3. Center for Research of Endemic Parasites of Iran (CREPI), Tehran University of Medical Sciences, Tehran, Iran
Department of Medical Parasitology and Mycology, School of Public Health
Iraj SHARIFI, Leishmaniasis Research Center, Dept. of Parasitology, Faculty of Medicine, Kerman University of Medical Sciences, Kerman, Iran
Leishmaniasis Research Center
Mohammadreza MAHMOUDI, Dept. of Microbiology and Parasitology, Faculty of Medicine, Guilan University of Medical Sciences, Rasht, Iran
Department of Microbiology and Parasitology, Faculty of Medicine
Asad MIRZAEI, Dept. of Parasitology, Faculty of Medicine, Ilam University of Medical Sciences, Ilam, Iran
Parasitology Department, Faculty of Medicine 
Mahmoodreza BEHRAVAN, Esfarayen Faculty of Medical Sciences, Esfarayen, Iran
Birjand Infection Disease Research Center & Microbiology Department
Mohsen ARBABI, Dept. of Parasitology and Mycology, School of Medicine, Kashan University of Medical Sciences, Kashan, Iran
Department of Medical Parasitology and Mycology, School of Medicine
Shahram NEKOEIAN, Dept. of Cellular and Molecular Biology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran
Department of Cellular and Molecular Biology 
Reza KALANTARI, Dept. of Parasitology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran
Department of Mycology & Parasitology, School of Medicine 
Behzad GHORBANZADEH, Dept. of Parasitology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran
Department of parasitology and entomology, faculty of medicine 

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Published
2018-09-23
How to Cite
1.
MOHAMMADIHA A, DALIMI A, MOHEBALI M, SHARIFI I, MAHMOUDI M, MIRZAEI A, SPOTIN A, BEHRAVAN M, KARIMI M, ARBABI M, NEKOEIAN S, KALANTARI R, GHORBANZADEH B. Molecular Identification and Phylogenetic Classification of Leishmania spp. Isolated from Human Cutaneous Leishmani-asis in Iran: A Cross-sectional Study. IJPA. 13(3):351-6.
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