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J. Clin. Microbiol. doi:10.1128/JCM.00236-07
Copyright (c) 2007, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Vibrio vulnificus typing based on Simple Sequence Repeats: insight to biotype 3

Department of Biotechnology and Food Engineering, Technion-Israel Institute of Technology, Haifa 32000, Israel; Government Central Laboratories, Ministry of Health, Jerusalem 94467, Israel; Department of Biology, Faculty of Sciences and Science Education, University of Haifa, Oranim, Tivon, 36006, Israel

^* To whom correspondence should be addressed. Email: kashi{at}techunix.technion.ac.il.

	Abstract

Vibrio vulnificus is an opportunistic highly invasive human pathogen with worldwide distribution. It is commonly divided into three biochemical groups (‘Biotypes’) most of which are pathogenic. Simple Sequence Repeats (SSR), provide a source of high genomic polymorphism used in bacterial typing. Here we describe the use of variation in mutable SSR loci for accurate and rapid genotyping of V. vulnificus. In-silico screen of the genome of two V. vulnificus strains revealed thousands of SSR tracts. Twelve SSR with core motif longer than 5 bp, were tested for polymorphism in a panel of 32 characterized and 56 other V. vulnificus isolates, representing both clinical and environmental isolates from all three biotypes. All tested SSR were polymorphic with diversity indices ranging from 0.17 to 0.90, revealing high discrimination among isolates (27 out of 32). Genetic analysis of the SSR data resulted in clear distinction of isolates that belong to the highly virulent biotype 3 group. Contrary to the clonal nature of this new group, SSR revealed high discriminatory power within biotype 3 group, as opposed to other molecular methods that failed to differentiate these isolates. Thus, SSRs are suitable for rapid typing and classification of V. vulnificus using high throughput capillary electrophoresis methods. SSR (5 bp) by their nature enable to identify variations occurring on a fine scale and therefore could provide new insights on the newly emerged biotype 3 group of V. vulnificus, and could be used as an efficient tool in epidemiological studies.