Japanese Journal of Food Microbiology
Online ISSN : 1882-5982
Print ISSN : 1340-8267
ISSN-L : 1340-8267
Methods
Development and Evaluation of the One-Shot Multiplex PCR Method Targeting Seven Major EHEC O Serogroups and Three Virulence Genes
Atsushi IGUCHIAtsuko AKIYOSHISunao IYODAMakoto OHNISHI
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JOURNAL FREE ACCESS

2015 Volume 32 Issue 4 Pages 215-218

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Abstract

Enterohemorrhagic E. coli (EHEC) or Shiga toxin-producing E. coli is known as an important food-borne pathogen, and O157 is the most frequently reported O serogroups of EHEC strains associated with bloody diarrhea and hemolytic-uremic syndrome worldwide. Subsequently O26, O111, O103, O145, O121 and O165 serogroups of EHEC were also frequently isolated from patients with severe diseases in Japan. The O serogrouping of EHEC is essential in outbreak investigations and surveillance. In a previous study, we developed a comprehensive and practical platform for molecular O-serogrouping of E. coli strains, named as E. coli O-genotyping PCR (Iguchi, A., et al.: J. Clin. Microbiol., in press), targeting unique sequences on each O-antigen biosynthesis gene cluster. Based on the PCR system, we now developed an EHEC-specific multiplex PCR system, named as “MP-1 plus,” targeting seven major EHEC O serogroups and three virulence genes, stx1, stx2 and eae. This primer set contains 10 primer pairs that amplify products with different sizes, and validation studies using reference and wild strains showed that the results were reliable enough. The one-shot PCR method reported here might be a promising tool for the identification and subtyping of EHEC strains for outbreak investigations as well as for the surveillance.

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© 2015 Japanese Society of Food Microbiology
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