Multiplex PCR Genotyping Assay That Distinguishes between Isolates ofClostridium perfringensType A Carrying a Chromosomal Enterotoxin Gene (cpe) Locus, a PlasmidcpeLocus with an IS1470-Like Sequence, or a PlasmidcpeLocus with an IS1151Sequence

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Abstract
Clostridium perfringenstype 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 chromosomalcpegene, while non-food-borne gastrointestinal (GI) disease isolates from those two geographic locations were found to have a plasmidcpegene. In this report, we describe the development of an economical multiplex PCRcpegenotyping assay that works with culture lysates to distinguish among type A isolates carrying a chromosomalcpegene, a plasmidcpegene with a downstream IS1470-like sequence, or a plasmidcpegene with a downstream IS1151sequence. When this multiplex PCR assay was applied in molecular epidemiologic studies, it was found that (i) all 57 examined type A isolates with a plasmidcpegene have either IS1470-like or IS1151sequences downstream of the plasmidcpegene; (ii) an IS1470-like sequence, rather than an IS1151sequence, is more commonly present downstream of the plasmidcpegene (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 chromosomalcpegene also appear to be the major cause ofC. perfringensfood poisoning in Japan. The superiority of this new multiplex PCR assay over existingcpegenotyping approaches should facilitate further molecular epidemiologic investigations ofC. perfringensenterotoxin-associated GI illnesses and their associatedcpe-positive type A isolates.

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