[Real-time PCR assay for rapid detection of Listeria monocytogenes in simulated milk specimens].

OBJECTIVE

To establish a real-time PCR assay for the rapid detection of Listeria monocytogenes in simulated milk specimens.

METHODS

Based on part fragments of hlyO gene, a pair of primers and Taq-Man probe were designed for quantitative detection of L. monocytogenes. The specificity of the primers and probe were tested by using different L. monocytogenes strains and other common pathogenic bacteria.

RESULTS

L. monocytogenes strains were positive in the detection and other tested strains were negative. The sensitivity of assay was 9 copies per PCR reaction.

CONCLUSION

The specificity and sensitivity of Taq Man real-time PCR technology for detecting L. monocytogenes in simulated dairy specimens were high, and the assay could be completed within 1.5 h. This method could be used to detect other food samples contaminated by L. monocytogenes and identify the cause of food-borne Listeriosis outbreaks.