title: Molecular Epidemiological Survey of Cutaneous Leishmaniasis in Two Highly Endemic Metropolises of Iran, Application of FTA Cards for DNA Extraction From Giemsa-Stained Slides.
abstract: BACKGROUND
  PCR has been used for confirmation of leishmaniasis in epidemiological studies, but complexity of DNA extraction and PCR approach has confined its routine use in developing countries.
  
  
  OBJECTIVE
  In this study, recent epidemiological situation of cutaneous leishmaniasis (CL) in two hyper-endemic metropolises of Shiraz and Isfahan in Iran was studied using DNA extraction by commercial FTA cards and kinetoplastid DNA (kDNA)-PCR amplification for detection/identification of Leishmania directly from stained skin scraping imprints.
  
  
  METHODS
  Fifty four and 30 samples were collected from clinically diagnosed CL patients referred to clinical laboratories of leishmaniasis control centers in Isfahan and Shiraz cities, respectively. The samples were examined by direct microscopy and then scrapings of the stained smears were applied to FTA cards and used directly as DNA source in a nested-PCR to amplify kDNA to detect and identify Leishmania species.
  
  
  RESULTS
  Fifty four of 84 (64.2%) slides obtained from patients had positive results microscopically, while 79/84 (94%) of slides had positive results by FTA card-nested-PCR. PCR and microscopy showed a sensitivity of 96.4% and 64.2% and specificity of 100% and 100%, respectively. Interestingly, Leishmania major as causative agent of zoonotic CL was identified in 100% and 90.7% of CL cases from Isfahan and Shiraz cities, respectively, but L. tropica was detected from only 9.3% of cases from Shiraz city. All cases from central regions of Shiraz were L. tropica and no CL case was found in Isfahan central areas.
  
  
  CONCLUSIONS
  Filter paper-based DNA extraction can facilitate routine use of PCR for diagnosis of CL in research and diagnostic laboratories in Iran and countries with similar conditions. Epidemiologic changes including dominancy of L. major in suburbs of Shiraz and Isfahan metropolises where anthroponotic CL caused by L. tropica had been established, showed necessity of precise studies on CL epidemiology in old urban and newly added districts in the suburbs.
authors.affiliation: Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, IR Iran.
authors.forename: Shahrokh
authors.initials: S
authors.lastname: Izadi
authors.affiliation: Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, IR Iran; Department of Medical Parasitology and Mycology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, IR Iran.
authors.forename: Hossein
authors.initials: H
authors.lastname: Mirhendi
authors.affiliation: National Health Research Center, Isfahan Health Research Center, Isfahan, IR Iran.
authors.forename: Niloufar
authors.initials: N
authors.lastname: Jalalizand
authors.affiliation: Department of Medical Parasitology and Mycology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, IR Iran.
authors.forename: Hossein
authors.initials: H
authors.lastname: Khodadadi
authors.affiliation: Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, IR Iran.
authors.forename: Mehdi
authors.initials: M
authors.lastname: Mohebali
authors.affiliation: Cellular and Molecular Department, Isfahan Province Health Center, Isfahan, IR Iran.
authors.forename: Shahram
authors.initials: S
authors.lastname: Nekoeian
authors.affiliation: Department of Medical Parasitology and Mycology, School of Medicine, Yasuj University of Medical Sciences, Yasuj, IR Iran.
authors.forename: Ali
authors.initials: A
authors.lastname: Jamshidi
authors.affiliation: Department of Medical Parasitology and Mycology, School of Medicine, Yasuj University of Medical Sciences, Yasuj, IR Iran; Cellular and Molecular Research Center, Yasuj University of Medical Sciences, Yasuj, IR Iran.
authors.forename: Mohammad Amin
authors.initials: MA
authors.lastname: Ghatee
keywords: Cutaneous leishmaniasis; Extraction Method; Iran; Molecular Epidemiology