Development of a multiplex PCR-based rapid typing method for enterohemorrhagic Escherichia coli O157 strains

Tadasuke Ooka, Jun Terajima, Masahiro Kusumoto, Atsushi Iguchi, Ken Kurokawa, Yoshitoshi Ogura, Md Asadulghani, Keisuke Nakayama, Kazunori Murase, Makoto Ohnishi, Sunao Iyoda, Haruo Watanabe, Tetsuya Hayashi

Research output: Contribution to journalArticlepeer-review

41 Citations (Scopus)

Abstract

Enterohemorrhagic Escherichia coli O157 (EHEC O157) is a food-borne pathogen that has raised worldwide public health concern. The development of simple and rapid strain-typing methods is crucial for the rapid detection and surveillance of EHEC O157 outbreaks. In the present study, we developed a multiplex PCR-based strain-typing method for EHEC O157, which is based on the variability in genomic location of IS629 among EHEC O157 strains. This method is very simple, in that the procedures are completed within 2 h, the analysis can be performed without the need for special equipment or techniques (requiring only conventional PCR and agarose gel electrophoresis systems), the results can easily be transformed into digital data, and the genes for the major virulence markers of EHEC O157 (the stx1, stx2, and eae genes) can be detected simultaneously. Using this method, 201 EHEC O157 strains showing different XbaI digestion patterns in pulsed-field gel electrophoresis (PFGE) analysis were classified into 127 types, and outbreak-related strains showed identical or highly similar banding patterns. Although this method is less discriminatory than PFGE, it may be useful as a primary screening tool for EHEC O157 outbreaks.

Original languageEnglish
Pages (from-to)2888-2894
Number of pages7
JournalJournal of Clinical Microbiology
Volume47
Issue number9
DOIs
Publication statusPublished - Sep 2009

All Science Journal Classification (ASJC) codes

  • Microbiology (medical)

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