Abstract:

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 Listeria monocytogenes and
Staphylococcus aureus in artificially inoculated meat.

Key words: TaqMan, hlyA gene of Listeria monocytogenes, nuc gene of Staphylococcus aureus, fluorescence quantitative polymerase chain reaction (PCR)