Genotyping of Enterotoxigenic Clostridium perfringens Fecal Isolates Associated with Antibiotic-Associated Diarrhea and Food Poisoning in North America

doi:10.1128/JCM.39.3.883-888.2001

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Journal of Clinical Microbiology, March 2001, p. 883-888, Vol. 39, No. 3
0095-1137/01/$04.00+0 DOI: 10.1128/JCM.39.3.883-888.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Genotyping of Enterotoxigenic Clostridium perfringens Fecal Isolates Associated with Antibiotic-Associated Diarrhea and Food Poisoning in North America

Shauna G. Sparks,¹ Robert J. Carman,² Mahfuzur R. Sarker,¹^, and Bruce A. McClane¹^,^*

Department of Molecular Genetics and Biochemistry, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15261,¹ and TechLab, Inc., Corporate Research Center, Blacksburg, Virginia 24060²

Received 29 September 2000/Returned for modification 28 November 2000/Accepted 8 December 2000

Clostridium perfringens type A isolates producing enterotoxin (CPE) are an important cause of food poisoning and non-food-bornehuman gastrointestinal (GI) diseases, including antibiotic-associateddiarrhea (AAD). Recent studies suggest that C. perfringens typeA food poisoning is caused by C. perfringens isolates carryinga chromosomal cpe gene, while CPE-associated non-food-borne GIdiseases, such as AAD, are caused by plasmid cpe isolates. Thoseputative relationships, obtained predominantly with European isolates,were tested in the current study by examining 34 cpe-positive,C. perfringens fecal isolates from North American cases of foodpoisoning or AAD. These North American disease isolates were allclassified as type A using a multiplex PCR assay. Furthermore,restriction fragment length polymorphism and pulsed-field gelelectrophoresis genotyping analyses showed the North AmericanAAD isolates included in this collection all have a plasmid cpegene, but the North American food poisoning isolates all carrya chromosomal cpe gene. Western blotting demonstrated CPE expressionby nearly all of these disease isolates, confirming their virulencepotential. These findings with North American isolates provideimportant new evidence that, regardless of geographic origin ordate of isolation, plasmid cpe isolates cause most CPE-associatedAAD cases and chromosomal cpe isolates cause most C. perfringenstype A food poisoning cases. These findings hold importance forthe development of assays for distinguishing cases of CPE-associatedfood-borne and non-food-borne human GI illnesses and also identifypotential epidemiologic tools for determining the reservoirs fortheseillnesses.

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

Present address: Department of Microbiology, Oregon State University, Corvallis, OR97331.

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