Ribotyping and assessment of toxigenic genes of clostridium difficile strains isolated from raw meat

Abstract

Background: Clostridium difficile is an anaerobic, spore forming, rod shaped bacterium that can produce toxins and may transmit through some types of food samples. The present survey was done to assess the prevalence, toxigenic gene profile and ribotyping of C. difficile strains isolated from raw bovine and ovine meat samples. Methods: Two hundred and fifty raw meat samples were collected from Isfahan, Iran and analyzed for presence of C. difficile using the microbial culture. Genomic DNA was extracted from C. difficile isolates. Extracted DNA samples were subjected to PCR for determination of tcdA, tcdB, and cdtA toxigenic genes. Ribotyping was done using the 16S and 23S rRNA genes amplification in a separated PCR assay. Results: Fifteen out of 250 (6%) raw meat samples were contaminated with C. difficile. Prevalence of C. difficile strains amongst the raw bovine and ovine meat samples were 8% and 4%, respectively. The most commonly detected toxigenic genes were tcdA (33.33%) and cdtA (20%). Distribution of tcdB was 13.33%. Amongst the combined toxigenic genes, tcdA+cdtA had the highest distribution (13.33%).