title: Leishmania major: genetic heterogeneity of Iranian isolates by single-strand conformation polymorphism and sequence analysis of ribosomal DNA internal transcribed spacer.
abstract: Protozoan parasites of Leishmania major are the causative agents of cutaneous leishmaniasis in different parts of Iran. We applied PCR-based methods to analyze L. major parasites isolated from patients with active lesions from different geographic areas in Iran in order to understand DNA polymorphisms within L. major species. Twenty-four isolates were identified as L. major by RFLP analysis of the ribosomal internal transcribed spacer 1 (ITS1) amplicons. These isolates were further studied by single-strand conformation polymorphism (SSCP) analysis and sequencing of ITS1 and ITS2. Data obtained from SSCP analysis of the ITS1 and ITS2 loci revealed three and four different patterns among all studied samples, respectively. Sequencing of ITS1 and ITS2 confirmed the results of SSCP analysis and showed the potential of the PCR-SSCP method for assessing genetic heterogeneity within L. major. Different patterns in ITS1 were due to substitution of one nucleotide, whereas in ITS2 the changes were defined by variation in the number of repeats in two polymorphic microsatellites. In total five genotypic groups LmA, LmB, LmC, LmD and LmE were identified among L. major isolates. The most frequent genotype, LmA, was detected in isolates collected from different endemic areas of cutaneous leishmaniasis in Iran. Genotypes LmC, LmD and LmE were found only in the new focus of CL in Damghan (Semnan province) and LmB was identified exclusively among isolates of Kashan focus (Isfahan province). The distribution of genetic polymorphisms suggests the existence of distinct endemic regions of L. major in Iran.
authors.affiliation: Ali-Ebne Abitaleb Hospital, Rafsanjan University of Medical Sciences, Rafsanjan, Iran. m_tashakori44@yahoo.com
authors.forename: Mahnaz
authors.initials: M
authors.lastname: Tashakori
authors.forename: Tashakori
authors.initials: T
authors.lastname: Mahnaz
authors.forename: Katrin
authors.initials: K
authors.lastname: Kuhls
authors.forename: Kuhls
authors.initials: K
authors.lastname: Katrin
authors.forename: Amer
authors.initials: A
authors.lastname: Al-Jawabreh
authors.forename: Al-Jawabreh
authors.initials: AJ
authors.lastname: Amer
authors.forename: Isabel L
authors.initials: IL
authors.lastname: Mauricio
authors.forename: Mauricio
authors.initials: M
authors.lastname: Isabel
authors.forename: Gabriele
authors.initials: G
authors.lastname: Schönian
authors.forename: Schönian
authors.initials: S
authors.lastname: Gabriele
authors.forename: Safar
authors.initials: S
authors.lastname: Farajnia
authors.forename: Farajnia
authors.initials: F
authors.lastname: Safar
authors.forename: Mohammad Hossein
authors.initials: MH
authors.lastname: Alimohammadian
authors.forename: Alimohammadian Mohammad
authors.initials: AM
authors.lastname: Hossein
mesh_terms: D000818:Animals; D021903:DNA, Ribosomal Spacer / Q000235:genetics*; D006801:Humans; D007492:Iran / Q000453:epidemiology; D018320:Leishmania major / Q000235:genetics* / Q000302:isolation & purification*; D016773:Leishmaniasis, Cutaneous / Q000453:epidemiology / Q000469:parasitology*; D018807:Polymorphism, Single-Stranded Conformational
chemical_list: D021903:DNA, Ribosomal Spacer