Genotyping of EnterotoxigenicClostridium perfringensFecal Isolates Associated with Antibiotic-Associated Diarrhea and Food Poisoning in North America

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Abstract
Clostridium perfringenstype A isolates producing enterotoxin (CPE) are an important cause of food poisoning and non-food-borne human gastrointestinal (GI) diseases, including antibiotic-associated diarrhea (AAD). Recent studies suggest thatC. perfringenstype A food poisoning is caused byC. perfringensisolates carrying a chromosomalcpegene, while CPE-associated non-food-borne GI diseases, such as AAD, are caused by plasmidcpeisolates. Those putative relationships, obtained predominantly with European isolates, were tested in the current study by examining 34cpe-positive,C. perfringensfecal isolates from North American cases of food poisoning or AAD. These North American disease isolates were all classified as type A using a multiplex PCR assay. Furthermore, restriction fragment length polymorphism and pulsed-field gel electrophoresis genotyping analyses showed the North American AAD isolates included in this collection all have a plasmidcpegene, but the North American food poisoning isolates all carry a chromosomalcpegene. Western blotting demonstrated CPE expression by nearly all of these disease isolates, confirming their virulence potential. These findings with North American isolates provide important new evidence that, regardless of geographic origin or date of isolation, plasmidcpeisolates cause most CPE-associated AAD cases and chromosomalcpeisolates cause mostC. perfringenstype A food poisoning cases. These findings hold importance for the development of assays for distinguishing cases of CPE-associated food-borne and non-food-borne human GI illnesses and also identify potential epidemiologic tools for determining the reservoirs for these illnesses.