DETECTION OF ENTEROTOXIGENIC CLOSTRIDIUM-PERFRINGENS IN FOOD AND FECAL SAMPLES WITH A DUPLEX PCR AND THE SLIDE LATEX AGGLUTINATION-TEST

A duplex PCR procedure was evaluated for the detection of Clostridium perfringens in food and biological samples and for the identification of enterotoxigenic strains, This method uses two sets of primers which amplify in the same reaction two different DNA fragments simultaneous ly: the 283-bp C, perfringens phospholipase C gene fragment. and the 4 26-bp enterotoxin gene fragment. Internal primers within the two prime r jets confirmed the specificity of the method by DNA-DNA hybridizatio n with the PCR products, No cross-reaction was observed with other Clo stridium species or with other bacteria routinely found in food, The d etection level was approximately 10(5) C, perfringens cells per g of s tool or food sample, when overnight enrichment culture was used, 10 C, perfringens cells per g was detected in 57 artificially contaminated food samples, The duplex PCR is a rapid, sensitive, and reliable metho d for the detection and identification of enterotoxigenic C. perfringe ns strains in food samples. A slide latex agglutination test was also evaluated as a rapid, simple technique for the detection of C, perfrin gens enterotoxin in stool samples.