Simultaneous detection of Carnobacterium and Leuconostoc in meat products by multiplex PCR.

摘要

A multiplex PCR approach for simultaneous detection of Leuconostoc and Carnobacterium and its validation in meat products is described. 2 multiplex PCR assays were developed using newly designed 16S rDNA-directed primers adapted to the current taxonomic situation of Leuconostoc and Carnobacterium that allowed: simultaneous detection of both genera and members of nonmotile species belonging to the genus Carnobacterium; and identification in a single assay of the nonmotile species C. divergens, C. maltaromicum and C. gallinarum. Sensitivities of 103 and 104 cfu/g were achieved for multiplex PCR detection of Carnobacterium and Leuconostoc, respectively, following artificially inoculated meat trials. Both multiplex PCR assays were also validated in 14 naturally contaminated samples covering 9 types of meat products. Results obtained by colony identification were confirmed using PCR. Methods described in this study provide a rapid and reliable tool for PCR detection of Carnobacterium and Leuconostoc in meat products, and for colony identification. This multiplex PCR approach will improve the analysis of spoilage microorganisms of refrigerated vacuum-packaged meat products in order to determine the most appropriate preservation method.