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Multiplex polymerase chain reaction (mPCR), the simultaneous amplification of two or more polymerase chain reaction (PCR) products in the same reaction tube, can be applied to the rapid detection of pathogenic microorganisms in food matrices. mPCR can be used to test for multiple pathogens simultaneously or to test for multiple virulence factors associated with a single pathogen. Heterogeneous pathogens such as Escherichia coli cannot only be detected by mPCR but can also be differentiated by pathotype or serotype based on amplicon profiles. Sensitive detection of pathogens in food by mPCR (≤1 cfu/g) can be achieved if a high concentration of target DNA with minimal contamination by PCR inhibitors can be prepared. This process generally requires enrichment, which adds 6–24 h to the assay time. Even so, pathogens that grow slowly, are difficult to isolate or are difficult to identify can be detected more rapidly by mPCR than by culture‐based methods. This review covered methods and application of mPCR assays for detection of pathogens in the food production and distribution process.

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