Synthetic DNA probes for detection of enterotoxigenic Clostridium perfringens strains isolated from outbreaks of food poisoning

Abstract

Four synthetic oligonucleotides encoding different parts of the Clostridium perfringens enterotoxin gene were used to test the enterotoxigenicity of C. perfringens strains isolated from confirmed outbreaks of food poisoning. Of the 245 strains isolated from food and feces originating from 186 separate outbreaks, 145 (59%) gave hybridization reactions with each of the four DNA probes used, while 104 strains did not hybridize with any of the probes. There was no correlation between serotype and the presence of the enterotoxin gene, although the C. perfiringens enterotoxin gene was rarely detected among nontypable strains (17%). Results show that DNA hybridization is a suitable method for the identification of C. perfringens strains which have the potential to produce enterotoxin.

Clostridial enteric diseases of domestic animals

“…In fact, an increasing body of evidence suggests a role for enterotoxigenic strains of type A in the etiology of diarrheal conditions in several animal species (110,292,293). Enterotoxigenic strains have been detected in many species of animals and in the environment (423)(424)(425). In one study, 6% of isolates from food handlers were enterotoxigenic, as were 2% from dogs, 12% from oysters, and 10% from water (361).…”

“…These methods can be especially useful in determining the ability or potential ability of an isolate to produce CPE, since they do not require maintenance of cell cultures for toxin assays, and isolates that have cpe but do not sporulate in vitro can be readily detected (reducing or eliminating false negatives). Several groups have reported methods for detection of cpe by use of both oligonucleotide probes (87,89,418,425) and PCR (114,115,207,220,360). As many as 22% of some species of domestic animals carried cpe-positive C. perfringens, as determined by colony hybridization methods (418).…”

“…As many as 22% of some species of domestic animals carried cpe-positive C. perfringens, as determined by colony hybridization methods (418). Six of 98 strains of C. perfringens from feces of farm animals contained cpe, based on screening with an oligonucleotide probe (424,425). Further study of 245 food and fecal isolates from nearly 200 outbreaks of food poisoning revealed that 59% were cpe positive by hybridization.…”

MEMBRANE FILTRATION IMMUNOSTAINING TECHNIQUE FOR DETECTION AND QUANTITATION OF ENTEROTOXIGENIC CLOSTRIDIUM PERFRINGENS IN PRECOOKED BEEF 1

“…While one of the limitations of the conventional plate count is that the minimum number of bacteria that can be enumerated is 10 CFUig and the agar plates require incubation for at least 18 h, the technique reported here is sensitive enough to detect as low as 2 CFU/g of meat samples within 6 h of incubation followed by 3% h staining procedure. The total detection time (about 10 h) from the processing and sampling of the meat of the final staining is comparable with gene-probe (Van Damme-Jongsten et al 1990) or polymerase chain reaction procedures (Saito et al 1992) where time consuming enrichments or sample extraction steps are usually required. The MFI procedure also has the added advantage of providing evidence for the presence of the enterotoxigenic C. perfringens in the analyzed sample, whereas the nucleic acid detection techniques (Van Damme-Jongsten et al 1990;Saito et al 1992) provide evidence for the presence of the gene but not for its product.…”

“…The total detection time (about 10 h) from the processing and sampling of the meat of the final staining is comparable with gene-probe (Van Damme-Jongsten et al 1990) or polymerase chain reaction procedures (Saito et al 1992) where time consuming enrichments or sample extraction steps are usually required. The MFI procedure also has the added advantage of providing evidence for the presence of the enterotoxigenic C. perfringens in the analyzed sample, whereas the nucleic acid detection techniques (Van Damme-Jongsten et al 1990;Saito et al 1992) provide evidence for the presence of the gene but not for its product. Compared with the commercial latex agglutination test (Harmon and Kautter 1986), the MFI procedure not only detects the enterotoxigenic C. perfringens, but it also provides an estimate of cell numbers in the food sample analyzed.…”