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Journal of Clinical Microbiology, April 2003, p. 1494-1498, Vol. 41, No. 4
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.4.1494-1498.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Development of a Duplex PCR Genotyping Assay for Distinguishing Clostridium perfringens Type A Isolates Carrying Chromosomal Enterotoxin (cpe) Genes from Those Carrying Plasmid-Borne Enterotoxin (cpe) Genes

Qiyi Wen,1,2 Kazuaki Miyamoto,1,3 and Bruce A. McClane1*

Department of Molecular Genetics and Biochemistry, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15261,1 Department of Veterinary Medicine, University of Yangzhou School of Veterinary Medicine, Yangzhou, Jiangsu 225009, China,2 Department of Microbiology, Wakayama Medical College, Wakayama, Japan 641-00213

Received 14 October 2002/ Returned for modification 9 December 2002/ Accepted 10 January 2003

About 5% of Clostridium perfringens type A isolates carry the cpe gene encoding the C. perfringens enterotoxin. Those cpe-positive type A isolates are important causes of food-poisoning and non-food-borne cases of diarrheas in humans, as well as certain veterinary cases of diarrhea. Previous studies have determined that the enterotoxigenic type A isolates causing both non-food-borne human gastrointestinal disease and veterinary disease carry their cpe genes on plasmids, while the type A isolates causing human food poisoning carry a chromosomal cpe gene. The present study reports on the successful development of a duplex PCR assay that can rapidly genotype enterotoxigenic type A isolates (i.e., determine whether those cpe-positive isolates carry a chromosomal or a plasmid-borne cpe gene). The availability of this rapid cpe genotyping assay capable of handling large numbers of samples provides a powerful new investigative tool for diagnostic, epidemiologic, and basic research purposes.

* Corresponding author. Mailing address: E1240 BST, Department of Molecular Genetics and Biochemistry, University of Pittsburgh School of Medicine, Pittsburgh, PA 15261. Phone: (412) 648-9022. Fax: (412) 624-1401. E-mail: bamcc{at}pitt.edu.

Journal of Clinical Microbiology, April 2003, p. 1494-1498, Vol. 41, No. 4
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.4.1494-1498.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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