Abstract: Objective: To develop a rapid multiplex polymerase chain reaction (m-PCR) assay for simultaneous detection of Salmonella spp., Staphylococcus aureus and Listeria monocytogenes in food. Methods: The genomic alignment method was used to identify specific PCR target sequences. Three pairs of specific primers were designed with Primer Premier 5.0 according to the invasive protein gene (invA) of Salmonella, the nuc gene of Staphylococcus aureus and the prs gene of Listeria monocytogen. Multiplex PCR was established by optimizing the reaction system. Results: The sensitivity of the multiplex PCR method was 7.6 pg/μL for Salmonellla spp., 3.8 pg/μL for Staphylococcus aureus, and 5.1 pg/μL for Listeria monocytogenes. All specific primers were amplified, and their corresponding strips were observed in validation experiments without cross reactivity. Conclusions: A triple PCR assay has been established for the simultaneous, sample, rapid and sensitive detection of Salmonella spp., Staphylococcus aureus and Listeria monocytogenes in food.

Key words: food-borne pathogenic bacteria, multiplex polymerase chain reaction (m-PCR), Salmonella spp., Staphylococcus aureus, Listeria monocytogen, food inspection