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Detection of enterotoxigenic Clostridium perfringens with a duplex PCR

E. AUGUSTYNOWICZ, A. GZYL and J. LUSARCZYK

Department of Sera and Vaccines Evaluation, National Institute of Hygiene, 24 Chocimska Street, 00-791 Warsaw, Poland

Corresponding author: Dr E. Augustynowicz (e-mail: eaugustynowicz{at}pzh.gov.pl).

Received 5 Jan. 2000; revised version accepted 27 July 2001.

Abstract

Two sets of primers designed to detect Clostridium perfringens phospholipase C (plc) and enterotoxin (cpe) genes in a single PCR reaction were applied to a collection of 64 predominantly food poisoning-related C. perfringens isolates. In-vitro enterotoxin synthesis was tested serologically after inducing sporulation. Of the 64 isolates, 26 were clearly enterotoxigenic; 16 were classified as potentially enterotoxigenic only as serological testing did not confirm enterotoxin production. Duplex PCR for diagnosis of enterotoxigenic C. perfringens from vegetative cultures can be a useful tool as fresh isolates often sporulate poorly or not all, giving rise to the possibility of false negative results by serological analysis.