Detection and characterisation of enterotoxigenic Clostridium perfringens type A isolates in Japanese retail meats
Abstract number: 1733_737
Miki Y., Kaneko-Hirano I., Miyamoto K., Fujiuchi K., Akimoto S.
Objectives:Clostridium perfringens is an important pathogen causing food-borne gastrointestinal disease. In Japan, C. perfringens is ranked as the third or forth greatest cause of the disease and sickened approximately 4,000 people, while the outbreak annually occurred 20 to 40 cases. Previous surveys found that the prevalence of enteropathogenic C. perfringens in Japanese retail food was approximately 04%, which was similar to that in USA. It is thought that food poisoning isolates carry cpe on their chromosome, while isolates from other gastrointestinal diseases, such as antibiotic-associated diarrhoea, carry cpe on transferable plasmid. In Japan, recently two food-borne outbreaks by plasmid-cpe isolates were reported, while several C. perfringens food poisoning isolates carry chromosomal cpe. In this survey, we investigated the prevalence of enterotoxigenic C. perfringens in Japanese retail meat samples, and then cpe-positive isolates were tested with multiplex PCR, which could differentiate chromosomal cpe isolate versus plasmid cpe isolate.
Methods: We collected two hundred meat samples from grocery stores and meat shops in Wakayama City. Approximately 100 g of food samples were anaerobically incubated at 45°C overnight with TGC II medium in stomacher bag, and then cultured samples were tested for the presence of cpe-positive C. perfringens followed by further incubation on SFP agar plate for bacterial isolation. DNA were prepared from overnight culture with InstaGene matrix (BioRad) and then used for PCR reaction for cpe detection.
Results: In PCR reaction, we detected cpe from 8 samples (4%), from three of which, we isolated cpe-positive C. perfringens. Interestingly, all three cpe-positive isolates carry cpe on the plasmid and it is combined with downstream IS1470-like structure in PCR cpe-genotyping assay.
Conclusion: From these results, C. perfringens carrying cpe on the plasmid might be a possible pathogen of C. perfringens food poisoning in Japan.
|Session name:||European Society of Clinical Microbiology and Infectious Diseases|
|Location:||ICC, Munich, Germany|
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