28
views
0
recommends
+1 Recommend
0 collections
    1
    shares
      • Record: found
      • Abstract: found
      • Article: found
      Is Open Access

      Characterization of Leptospira interrogans Serovars by Polymorphism Variable Number Tandem Repeat Analysis

      research-article

      Read this article at

      Bookmark
          There is no author summary for this article yet. Authors can add summaries to their articles on ScienceOpen to make them more accessible to a non-specialist audience.

          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.

          Objectives:

          The propose of this study was genomic identification of pathogenic Leptospires in Iran by MLVA.

          Materials and 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.

          Related collections

          Most cited references22

          • Record: found
          • Abstract: found
          • Article: not found

          Application of multilocus variable-number tandem-repeat analysis for molecular typing of the agent of leptospirosis.

          Leptospirosis is a worldwide-distributed zoonosis, endemic in tropical areas. Epidemiologic investigations of leptospirosis still rely on tedious serological identification tests. Recently, molecular typing systems based on variable-number tandem-repeat (VNTR) analysis have been described and have been used to identify Leptospira interrogans strains. Although L. interrogans is the most common Leptospira species encountered in human infections around the world, other pathogenic species, such as Leptospira kirschneri and Leptospira borgpetersenii, are also frequently associated with human leptospirosis. In this study, we aimed to extend multilocus VNTR analysis (MLVA) identification of strains to species other than L. interrogans. We designed primers for VNTR loci found in L. interrogans, L. kirschneri, and L. borgpetersenii. The discriminatory power of the redefined primers was evaluated on collection strains and then on clinical strains. We also carried out a retrospective study on 156 strains isolated from patients and animals from New Caledonia, an area of high endemicity in the South Pacific. Our results show that this simple PCR-based MLVA typing technique is a powerful methodology for the epidemiology of leptospirosis.
            Bookmark
            • Record: found
            • Abstract: found
            • Article: not found

            Identification of novel intergenic repetitive units in a mycobacterial two-component system operon.

            Mycobacterial interspersed repetitive units (MIRUs), a novel class of repeated sequences, were identified within the intercistronic region of an operon coding for a mycobacterial two-component system, named senX3-regX3. Southern blot analysis and homology searches revealed the presence of several homologous sequences in intergenic regions dispersed throughout the genomes of Mycobacterium bovis BCG, Mycobacterium tuberculosis and Mycobacterium leprae. These could be grouped into three major families, containing elements of 77-101 bp, 46-53 bp and 58-101 bp. Based on the available mycobacterial sequences, the total number of MIRUs is estimated to be about 40-50 per genome. Similar to previously identified small repetitive sequences, the MIRUs of the two-component operon are transcribed on a polycistronic mRNA. Unlike previously identified small repetitive sequences, however, MIRUs do not contain dyad symmetries, comprise small open reading frames (ORFs) whose extremities overlap those of the contiguous ORFs and are oriented in the same translational direction as those of the adjacent genes. Analyses of the sequences at the insertion sites suggest that MIRUs disseminate by transposition into DTGA sites involved in translational coupling in polycistronic operons.
              Bookmark
              • Record: found
              • Abstract: found
              • Article: not found

              Identification of variable-number tandem-repeat loci in Leptospira interrogans sensu stricto.

              Leptospira interrogans sensu stricto is responsible for the most frequent and severe cases of human leptospirosis. The epidemiology and clinical features of leptospirosis are usually associated with the serovars and serogroups of Leptospira. Because of the difficulties associated with serological identification of Leptospira strains, we evaluated a novel PCR-based method for typing L. interrogans serovars. Based upon the genome sequence of L. interrogans serovar Lai type strain 5660, 44 loci were analyzed by PCR for their variability in size due to the presence of variable-number tandem repeats (VNTR). Seven VNTR loci were found to be powerful markers for serovar identification, epidemiology, and phylogenetic studies of L. interrogans. This rapid and easy method should greatly contribute to a better knowledge of the epidemiology of Leptospira.
                Bookmark

                Author and article information

                Journal
                Jundishapur J Microbiol
                Jundishapur J Microbiol
                10.5812/jjm
                Kowsar
                Jundishapur Journal of Microbiology
                Kowsar
                2008-3645
                2008-4161
                18 October 2015
                October 2015
                : 8
                : 10
                : e22819
                Affiliations
                [1 ]Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, IR Iran
                [2 ]Department of Medical Microbiology, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, IR Iran
                [3 ]Department of Microbiology, Razi Vaccine and Serum Institute, Karaj, IR Iran
                [4 ]Department of Celiac Disease, Shahid Beheshti University of Medical Sciences, Tehran, IR Iran
                [5 ]Department of Microbiology, Faculty of Basic Sciences, Karaj Branch, Islamic Azad University, Karaj, IR Iran
                [6 ]Department of Microbiology, Faculty of Basic Sciences, Zanjan Branch, Islamic Azad University, Zanjan, IR Iran
                Author notes
                [* ]Corresponding author: Sama Rezasoltani, Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, IR Iran. Tel: +98-9123197258, Fax: +98-2188656198, E-mail: samasoltani70@ 123456gmail.com
                Article
                10.5812/jjm.22819
                4641467
                4ecc65b7-d807-46f2-8ee1-00238327b78c
                Copyright © 2015, Ahvaz Jundishapur University of Medical Sciences.

                This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International License ( http://creativecommons.org/licenses/by-nc/4.0/) which permits copy and redistribute the material just in noncommercial usages, provided the original work is properly cited.

                History
                : 16 August 2014
                : 23 January 2015
                : 16 February 2015
                Categories
                Research Article

                mlva technique,vntr analysis,polymorphism,leptospira serovars

                Comments

                Comment on this article