Development of Target-Enriched Multiplex PCR Assay for Simultaneous Detection of Three Foodborne Pathogens

In this study,a target-enriched multiplex PCR(Tem-PCR) assay for simultaneous detection of Salmonella,Staphylococcus aureus and Shigella was developed. Based on the S. aureus fem A gene,the Salmonella inv A gene and the Shigella ipa H gene,three pairs of specific primers together with a universal primer were designed. Following the optimization of reaction conditions,the Tem-PCR assay was successfu lly established. Results showed that the TemPCR assay was highly specific to the target bacteria with a sensitivity of 1.1×10~3 CFU/mL,2×10~3 CFU/mL and 1.2×10~3 CFU/mL for Salmonella,S. aureus and Shigella,respectively. The Tem-PCR assay can effectively improve the balanc e of amplification efficiencies of the primers e mployed in traditional multiplex PCR.