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Applied and Environmental Microbiology, October 2004, p. 6309-6314, Vol. 70, No. 10
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.10.6309-6314.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

SHORT REPORT

Genetic Typing of Shiga Toxin 2 Variants of Escherichia coli by PCR-Restriction Fragment Length Polymorphism Analysis

Liesbet De Baets,1,2* Imme Van der Taelen,1,2,{dagger} Marina De Filette,1,2,{ddagger} Denis Piérard,3 Lesley Allison,4 Henri De Greve,2,§ Jean-Pierre Hernalsteens,2 and Hein Imberechts1

Dienst Algemene Bacteriologie, Centrum voor Onderzoek in Diergeneeskunde en Agrochemie,1 Onderzoeksgroep Genetische Virologie, Vrije Universiteit Brussel,2 Afdeling Microbiologie, Academisch Ziekenhuis Vrije Universiteit Brussel, Brussels, Belgium,3 Scottish E. coli O157 Reference Laboratory, Department of Clinical Microbiology, Western General Hospital, Edinburgh, Scotland4

Received 20 November 2003/ Accepted 15 May 2004

ABSTRACT

Shiga toxins Stx1 and Stx2 play a prominent role in the pathogenesis of Shiga toxin-producing Escherichia coli (STEC) infections. Several variants of the stx2 gene, encoding Stx2, have been described. In this study, we developed a PCR-restriction fragment length polymorphism system for typing stx2 genes of STEC strains. The typing system discriminates eight described variants and allows the identification of new stx2 variants and STEC isolates carrying multiple stx2 genes. A phylogenetic tree, based on the nucleotide sequences of the toxin-encoding genes, demonstrates that stx2 sequences with the same PvuII HaeIII HincII AccI type generally cluster together.

* Corresponding author. Mailing address: Dienst Algemene Bacteriologie, CODA, Groeselenberg 99, B-1180 Ukkel, Belgium. Phone: 32-2-3790427. Fax: 32-2-3790670. E-mail: lideb{at}var.fgov.be.

FOOTNOTES

{dagger} Present address: Purna Pharmaceuticals NV/SA, B-2870 Puurs, Belgium.

{ddagger} Present address: Dienst Moleculaire Biologie, Universiteit Gent, B-9052 Zwijnaarde, Belgium.

§ Present address: Onderzoeksgroep Ultrastructuur, Vlaams Interuniversitair Instituut voor Biotechnologie, Vrije Universiteit Brussel, B-1050 Brussels, Belgium.

Applied and Environmental Microbiology, October 2004, p. 6309-6314, Vol. 70, No. 10
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.10.6309-6314.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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