Dual-priming primers-based PCR method for specific detection of Listeria monocytogenes

In this study,a Dual-priming oligonucleotide(DPO)-based PCR method for specific detection of L.monocytogenes was developed with iap gene as target. Annealing temperature insensitivity,specificity and detection sensitivity of the DPO-PCR method were tested and its preliminary application was carried out in practice. Results showed that the detection sensitivity of the DPO-PCR method was 151CFU / mL. In the annealing temperature insensitivity test,compared with conventional PCR primers,DPO primers were able to efficiently amplify target gene in the annealing temperature range of 48~68℃. In the specificity test,the DPOPCR method showed a higher specificity for the target bacteria than conventional PCR method and no nonspecific amplification reactions were observed in DPO-PCR. In practice,9 L.monocytogenes positive samples from 130 samples were detected by the DPO-PCR method,which was in accordance with the testing results according to GB/T 4789.30-2008,showing a better practicability. The DPO-PCR provided a new method for fast and accurate detection of L.monocytogenes.