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Journal of Clinical Microbiology, April 2004, p. 1552-1558, Vol. 42, No. 4
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.4.1552-1558.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Multiplex PCR Genotyping Assay That Distinguishes between Isolates of Clostridium perfringens Type A Carrying a Chromosomal Enterotoxin Gene (cpe) Locus, a Plasmid cpe Locus with an IS1470-Like Sequence, or a Plasmid cpe Locus with an IS1151 Sequence

Department of Microbiology, Wakayama Medical College, Wakayama, Japan 641-0021,¹ Department of Molecular Genetics and Biochemistry, School of Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania 15261,² School of Veterinary Medicine, University of Yangzhou, Yangzhou, Jiangsu 225009, China³

Received 20 September 2003/ Returned for modification 9 December 2003/ Accepted 7 January 2004

Clostridium perfringens type A isolates carrying the enterotoxin (cpe) gene are important causes of both food poisoning and non-food-borne diarrheas in humans. In North America and Europe, food poisoning isolates were previously shown to carry a chromosomal cpe gene, while non-food-borne gastrointestinal (GI) disease isolates from those two geographic locations were found to have a plasmid cpe gene. In this report, we describe the development of an economical multiplex PCR cpe genotyping assay that works with culture lysates to distinguish among type A isolates carrying a chromosomal cpe gene, a plasmid cpe gene with a downstream IS1470-like sequence, or a plasmid cpe gene with a downstream IS1151 sequence. When this multiplex PCR assay was applied in molecular epidemiologic studies, it was found that (i) all 57 examined type A isolates with a plasmid cpe gene have either IS1470-like or IS1151 sequences downstream of the plasmid cpe gene; (ii) an IS1470-like sequence, rather than an IS1151 sequence, is more commonly present downstream of the plasmid cpe gene (particularly in North American non-food-borne human GI disease isolates); and (iii) as previously shown in the United States and Europe, isolates carrying the chromosomal cpe gene also appear to be the major cause of C. perfringens food poisoning in Japan. The superiority of this new multiplex PCR assay over existing cpe genotyping approaches should facilitate further molecular epidemiologic investigations of C. perfringens enterotoxin-associated GI illnesses and their associated cpe-positive type A isolates.

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• Miki, Y., Miyamoto, K., Kaneko-Hirano, I., Fujiuchi, K., Akimoto, S. (2008). Prevalence and Characterization of Enterotoxin Gene-Carrying Clostridium perfringens Isolates from Retail Meat Products in Japan. Appl. Environ. Microbiol. 74: 5366-5372 [Abstract] [Full Text]  
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