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Journal of Clinical Microbiology, September 2007, p. 2951-2959, Vol. 45, No. 9
0095-1137/07/$08.00+0     doi:10.1128/JCM.00236-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Vibrio vulnificus Typing Based on Simple Sequence Repeats: Insights into the Biotype 3 Group ^,

Yoav Y. Broza,¹ Yael Danin-Poleg,¹ Larisa Lerner,² Meir Broza,³ and Yechezkel Kashi^1*

Department of Biotechnology and Food Engineering, Technion-Israel Institute of Technology, Haifa 32000,¹ Government Central Laboratories, Ministry of Health, Jerusalem 94467,² Department of Biology, Faculty of Sciences and Science Education, University of Haifa, Oranim, Tivon 36006, Israel³

Received 30 January 2007/ Returned for modification 18 March 2007/ Accepted 16 July 2007

Vibrio vulnificus is an opportunistic, highly invasive human pathogen with worldwide distribution. V. vulnificus strains are commonly divided into three biochemical groups (biotypes), most members of which are pathogenic. Simple sequence repeats (SSR) provide a source of high-level genomic polymorphism used in bacterial typing. Here, we describe the use of variations in mutable SSR loci for accurate and rapid genotyping of V. vulnificus. An in silico screen of the genomes of two V. vulnificus strains revealed thousands of SSR tracts. Twelve SSR with core motifs longer than 5 bp in a panel of 32 characterized and 56 other V. vulnificus isolates, including both clinical and environmental isolates from all three biotypes, were tested for polymorphism. All tested SSR were polymorphic, and diversity indices ranged from 0.17 to 0.90, allowing a high degree of discrimination among isolates (27 of 32 characterized isolates). Genetic analysis of the SSR data resulted in the clear distinction of isolates that belong to the highly virulent biotype 3 group. Despite the clonal nature of this new group, SSR analysis demonstrated high-level discriminatory power within the biotype 3 group, as opposed to other molecular methods that failed to differentiate these isolates. Thus, SSR are suitable for rapid typing and classification of V. vulnificus strains by high-throughput capillary electrophoresis methods. SSR (5 bp) by their nature enable the identification of variations occurring on a small scale and, therefore, may provide new insights into the newly emerged biotype 3 group of V. vulnificus and may be used as an efficient tool in epidemiological studies.

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* Corresponding author. Mailing address: Department of Biotechnology and Food Engineering, The Technion Haifa, Haifa 32000, Israel. Phone: 972-4-8293074. Fax: 972-4-8293399. E-mail: kashi{at}techunix.technion.ac.il

Published ahead of print on 25 July 2007.

Supplemental material for this article may be found at http://jcm.asm.org/.

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Journal of Clinical Microbiology, September 2007, p. 2951-2959, Vol. 45, No. 9
0095-1137/07/$08.00+0     doi:10.1128/JCM.00236-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

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