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Euroroundups Use of multilocus variable-number tandem repeat analysis (MLVA) in eight European countries, 2012 B A Lindstedt (bjorn-arne.lindstedt@fhi.no) 1 , M Torpdahl 2 , G Vergnaud 3,4 , S Le Hello 5 , F X Weill 5 , E Tietze 6 , B Malorny 7 , D M Prendergast 8 , E Ní Ghallchóir 8 , R F Lista 9 , L M Schouls 10 , R Söderlund 11 , S Börjesson 11 , S Åkerström 11 1. Division of Infectious Diseases Control, Norwegian Institute of Public Health, Oslo, Norway 2. Department of Microbiological Surveillance and Research, Statens Serum Institut, Copenhagen, Denmark 3. Université Paris-Sud, Institut de Génétique et Microbiologie, Unités Mixtes de Recherche (UMR) 8621, Orsay, France 4. Direction Générale de l’Armement (DGA)/Mission pour la Recherche et l’Innovation Scientifique (MRIS), Bagneux, France 5. Institut Pasteur, Unité de Recherche et d’Expertise des Bactéries Pathogènes Entériques, Centre National de Référence E. coli/Shigella/Salmonella, Paris, France 6. National Reference Center for Salmonella and other Enterics, Robert Koch Institute, Wernigerode Branch, Wernigerode, Germany 7. Federal Institute for Risk Assessment (BfR) National Salmonella Reference Laboratory Department Biological Safety, Berlin, Germany 8. Central Veterinary Research Laboratory, Department of Agriculture, Food and the Marine, Kildare, Ireland 9. Health Corps Italian Army, Department of Molecular Biology, Immunology and Experimental Medicine, Army Medical and Veterinary Research Center, Rome, Italy 10. Laboratory for Infectious Diseases and Perinatal Screening (LIS), Centre for Infectious Disease Control Netherlands (CIb), National Institute for Public Health and the Environment (RIVM), Bilthoven, the Netherlands 11. The National Veterinary Institute (SVA), Uppsala, Sweden Citation style for this article: Lindstedt BA, Torpdahl M, Vergnaud G, Le Hello S, Weill FX, Tietze E, Malorny B, Prendergast DM, Ní Ghallchóir E, Lista RF, Schouls LM, Söderlund R, Börjesson S, Åkerström S. Use of multilocus variable-number tandem repeat analysis (MLVA) in eight European countries, 2012. Euro Surveill. 2013;18(4):pii=20385. Available online: http://www.eurosurveillance.org/ViewArticle.aspx?ArticleId=20385 Genotyping of important medical or veterinary prokaryotes has become a very important tool during the last decades. Rapid development of fragment-separation and sequencing technologies has made many new genotyping strategies possible. Among these new methods is multilocus variable-number tandem repeat analysis (MLVA). Here we present an update on the use of MLVA in eight European countries (Denmark, France, Germany, Ireland, Italy, the Netherlands, Norway and Sweden). Researchers in Europe have been active in developing and implementing a large array of different assays. MLVA has been used as a typing tool in several contexts, from aiding in resolving outbreaks of foodborne bacteria to typing organisms that may pose a bioterrorist threat, as well as in scientific studies. Introduction Multilocus variable-number tandem repeat analysis (MLVA) is a DNA-based molecular typing method frequently applied to the study of prokaryotes. It records size polymorphisms in several variable-number of tandem repeats (VNTR) loci amplified by stringent PCR protocols. MLVA will mainly impact the public health field by introducing newer, faster and safer (reduced handling of live bacteria) methodologies for typing microorganisms. Reduced typing time, with high resolution, is beneficial for resolving large and complex outbreak situations. The methodology is also suitable for large-scale automation: suitable instruments (e.g. automated sequencers, pipetting robots and analytical software) are already commercially available. There are www.eurosurveillance.org Article submitted on 30 June 2012 / published on 24 January 2013 several variations of MLVA assays depending on available instrumentation. Earlier versions tended to measure VNTR sizes by agarose gel electrophoresis, while newer assays often use capillary electrophoresis for size determination once the allele size range at each locus has been well characterised. As mentioned above, MLVA assays have clear advantages, offering fast typing, high resolution and reduced handling times of pathogenic organisms. Their drawbacks include high assay-specificity (e.g. each organism usually needs a distinct MLVA assay) and the, as yet, lack of standardisation for the majority of published assays. In Europe, only the Salmonella enterica subspecies enterica serovar Typhimurium (S. Typhimurium) MLVA assay has achieved generally accepted standardisation [1,2]. MLVA is gaining in popularity: in 2000, there was only one PubMed entry (when searching for ‘MLVA’) while in 2011, there were 96 entries for articles that year alone. There has been extensive research on MLVA and MLVA protocol development within Europe: an overview of organisms for which there are existing MLVA assays in European countries, based on web searches for protocols is presented in Table 1. The web searches were performed on 23 April 2012 and repeated on 18 June in PubMed using the search terms; ‘MLVA’, ‘VNTR’, ‘tandem repeats’, ‘TR’, ‘direct repeats’, ‘DR’ and ‘genotyping’, combined with geographical names such as ‘Europe’, ‘European’ or the countries within Europe. General Internet searches using the same keywords in a standard web browser were also included. The same 39
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