Abstract:

A high-throughput and rapid multiplex PCR-denaturing high performance liquid chromatographic (MPCRDHPLC) method was presented for the detection of Listeria monocytogenes, Listeria ivanovii and Listeria innocua in food. Respective primers specific to target genes of Listeria monocytogenes, Listeria ivanovii and Listeria innocua were designed and synthesized, followed by multiplex PCR amplification and DHPLC analysis. Along with repeatability experiments, specificity experiments were performed using 94 reference strains. The MPCR-DHPLC method resulted in a simultaneous detection of specific positive absorption of Listeria monocytogenes, Listeria ivanovii and Listeria innocua rather than other relative species or reference strains of non-relative species and exhibited good specificity and repeatability and other advantages such as rapidity, accuracy and high throughput, thereby providing a novel technique for the detection of microorganisms in food.

Key words: MPCR-DHPLC, Listeria monocytogenes, Listeria ivanovii, Listeria innocua, detection