Detection of Clostridium perfringens and Bacillus cereus on Sus scrofa domesticus by Multiplex PCR

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Abstract Clostridium perfringens and Bacillus cereus are major foodborne pathogens associated with gastrointestinal diseases in humans and animals and are frequently detected in pork products. In Vietnam, where pork represents a major component of daily meat consumption, the presence of these pathogens poses significant food safety concerns. Conventional detection methods are often time-consuming, and unsuitable for rapid large-scale screening. This study aimed to develop a rapid and reliable multiplex PCR assay for the simultaneous detection of C. perfringens and B. cereus by targeting the species-specific SDR and DHDPS genes, respectively. The assay was evaluated using pure cultures, mixed bacterial samples, and swine intestinal specimens to assess its specificity and practical applicability. The multiplex PCR successfully generated distinct amplicons of 615 bp for C. perfringens and 218 bp for B. cereus , demonstrating high specificity without cross-reactivity. The method also effectively detected single and co-contamination in intestinal samples. These findings highlight the potential application of multiplex PCR as a rapid screening tool for improving pork safety surveillance and foodborne pathogen monitoring.

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