Prevalence of the Enterotoxin Gene and Clonality of Clostridium perfringens Strains Associated with Food-Poisoning Outbreaks

Abstract

The prevalence of the enterotoxin gene in a well-characterized collection of 71 Clostridium perfringens strains from 36 separate food-poisoning cases or outbreaks was analyzed with the polymerase chain reaction (PCR). The clonality of 39 strains originating from 14 outbreaks where at least two isolates were available was studied with pulsed-field gel electrophoresis (PFGE) using SmaI and ApaI restriction endonucleases. The cpe gene PCR assay was found to correlate well with Clostridium perfringens enterotoxin (CPE) production in vitro with reverse passive latex agglutination. Of the C. perfringens food and clinical food-poisoning isolates 24 (86%) and 38 (88%) were cpe-positive, respectively. Different PFGE patterns indicated that multiple cpe-positive clones are frequently present within one outbreak. The existence of cpe-positive and negative isolates with identical or nearly identical PFGE patterns in a single outbreak suggests that the cpe gene may be in a movable genetic element.

Enumeration and Isolation of cpe -Positive Clostridium perfringens Spores from Feces

“…The HGMF-CH method provides a new approach in the diagnostics of food poisoning caused by cpe-positive C. perfringens, since the conventional isolation of cpe-positive C. perfringens from the feces of affected individuals is occasionally complicated by the presence of cpe-negative C. perfringens (5,11,16,20,22,26). As the HGMF-CH method provides a means to determine the presence of the cpe gene in a large number of isolates simultaneously and to enumerate and further isolate the cpe-positive strain, the method markedly improves the investigation of the epidemiology of cpe-positive C. perfringens strains.…”

“…Several probe-positive colonies were picked from the original plates, streaked onto blood agar, and incubated for 24 h at 37°C. These isolates were further analyzed for the presence of the cpe gene by PCR (20).…”

“…When confirming a food poisoning outbreak caused by C. perfringens, isolation of the same cpe-positive strain from suspect food and from stools of affected individuals is advisable. However, conventional isolation of cpe-positive C. perfringens from the feces of affected individuals is occasionally complicated by the presence of cpenegative C. perfringens as part of the normal fecal microbial population (5,11,16,20,22,26). The isolation of the causative agent from fecal samples by direct plating is challenging, particularly if several days have passed since the onset of the illness (16).…”

Clostridium perfringensin poultry: an emerging threat for animal and public health

“…C. perfringens type A strains produce the chromosomal-encoded alpha-toxin, while C. perfringens type C strains produce alpha toxin together with beta toxin (Petit et al, 1999). Some strains of C. perfringens type A produce an enterotoxin at the moment of sporulation, causing disease in humans (Ridell et al, 1998;Sarker et al, 1999). The mechanism of action of the enterotoxin will be discussed further in the text.…”

“…The effects of type A food poisoning are caused by production of the Clostridium perfringens enterotoxin (CPE), which is produced at the moment of sporulation (Ridell et al, 1998;Sarker et al, 1999). The CPE appears to be a bifunctional toxin, which first induces cellular damage via its cytotoxic activity, resulting from plasma membrane permeability alterations caused by formation of CPE-containing complex that may correspond to a pore.…”