A Duplex Fluorescence Quantitative PCR Assay for Detecting Listeriamonocytogenes and Staphylococcus aureus

A TaqMan duplex fluorescence quantitative PCR assay for detecting Listeria monocytogenes and Staphylococcus aureus was presented.Specific primers and probes were designed according to the hlyA gene sequence of Listeria monocytogenes and the nuc gene of Staphylococcus aureus published in GenBank.The developed assays detected only the two pathogens,respectively,but not other bacteria,indicating the high specificity of the duplex PCR.When the positive recombinant plasmids pMD18-hlyA and pMD18-nuc were simultaneously amplified,the sensitivity for them was 19.5 copy/μL and 18.7 copy/μL,respectively,and the limit of detection was 102 CFU/g for both Listeriamonocytogenes and Staphylococcus aureus in artificially inoculated meat.