The Bacillus cereus sensu lato group represents a significant food safety concern, as its widespread presence in food, the environment and in the human gut contributes to frequent diarrheal and emetic foodborne illnesses. Despite the importance of its major virulence determinants, enterotoxins Hbl, Nhe, CytK, molecular characterization of toxin genes in Slovenian isolates remains limited. The aim of this study was to develop and optimize a realtime PCR method for the simultaneous detection of hblD, nheA and cytK2, and to assess the prevalence of these genes in isolates obtained from food, surface swabs and human feces samples. The method was optimized and validated stepwise. We examined 179 isolates in the study, among which the nheA gene was the most common gene (98 %), while the combination of all three diarrheal genes occured in approximately half of the isolates. A comparison of PCR with BCET RPLA results demonstrated high concordance (98 %). Overall, our findings show that the developed method provides a reliable genotypic tool for characterization of the enterotoxin profiles of strains from B. cereus s. l. group and reveal a broad distribution of toxin genes of this bacterial group across the studied environments.
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