title: Characterization of Leptospira interrogans Serovars by Polymorphism Variable Number Tandem Repeat Analysis.
abstract: BACKGROUND
  Leptospirosis is recognized as a re-emerging infectious disease; therefore, understanding the epidemiology of the disease is vital for designing intervention programs and diminishing its transmission. Recently, Multilocus variable number tandem repeat analysis (MLVA) is used for segregating and identifying Leptospira serovars. The method has potential application in investigating the molecular epidemiology of Leptospira.
  
  
  OBJECTIVE
  The propose of this study was genomic identification of pathogenic Leptospires in Iran by MLVA.
  
  
  METHODS
  Leptospira serovars were obtained from National Reference Laboratory of Leptospira at Razi Vaccine and Serum Research Institute, Karaj, Iran. Serovars were cultured into the liquid EMJH medium and incubated at 28˚C for 7 days. DNA of serovars was extracted using the phenol-chloroform method. PCR was performed with 5 selected variable number tandem repeat analysis (VNTR) loci. The amplified products were analyzed by agarose gel electrophoresis. The size of the amplified products was estimated by 100 bp ladder and sequencing analysis.
  
  
  RESULTS
  The saprophytic serovar showed no amplified fragments. PCR products in all pathogenic serovars were observed. The 12 reference serovars used for the development of technique displayed distinct patterns.
  
  
  CONCLUSIONS
  Results showed that MLVA technique with its range of polymorphism is a good marker for identification of pathogenic serovars. Some VNTR loci are more powerful than the other ones with regard to differentiation. Serovars from the same geographical area have more genetic similarity than same serovars from different places. MLVA is a suitable technique for epidemiological survey.
authors.affiliation: Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, IR Iran.
authors.forename: Sama
authors.initials: S
authors.lastname: Rezasoltani
authors.affiliation: Department of Medical Microbiology, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, IR Iran.
authors.forename: Hossein
authors.initials: H
authors.lastname: Dabiri
authors.affiliation: Department of Microbiology, Razi Vaccine and Serum Institute, Karaj, IR Iran.
authors.forename: Pejvak
authors.initials: P
authors.lastname: Khaki
authors.affiliation: Department of Celiac Disease, Shahid Beheshti University of Medical Sciences, Tehran, IR Iran.
authors.forename: Mohammad
authors.initials: M
authors.lastname: Rostami Nejad
authors.affiliation: Department of Microbiology, Faculty of Basic Sciences, Karaj Branch, Islamic Azad University, Karaj, IR Iran.
authors.forename: Nasim
authors.initials: N
authors.lastname: Karimnasab
authors.affiliation: Department of Microbiology, Faculty of Basic Sciences, Zanjan Branch, Islamic Azad University, Zanjan, IR Iran.
authors.forename: Shiva
authors.initials: S
authors.lastname: Modirrousta
keywords: Leptospira Serovars; MLVA Technique; Polymorphism; VNTR Analysis