Simultaneous detection of Carnobacterium and Leuconostoc in meat products by multiplex PCR.
OBJECTIVE
To develop a multiplex PCR approach for simultaneous detection of Leuconostoc and Carnobacterium and its validation in meat products.


RESULTS
Two multiplex PCR assays were developed using newly designed 16S rDNA-directed primers adapted to the current taxonomic situation of genera Leuconostoc and Carnobacterium that allow: (i) simultaneous detection of both genera, and members of the nonmotile species of genus Carnobacterium and (ii) identification in a single assay of the nonmotile species C. divergens, C. maltaromicum and C. gallinarum. Sensitivity values of 10(3) and 10(4) CFU g(-1) were determined for multiplex PCR detection of Carnobacterium and Leuconostoc, respectively, following artificially inoculated meat trials. In addition, both multiplex PCR assays were validated in 14 naturally contaminated samples covering nine types of meat products. Results obtained by colony identification were confirmed by PCR detection.


CONCLUSIONS
The 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.


CONCLUSIONS
This multiplex PCR approach will help in the analysis of the spoilage microbiota of refrigerated vacuum-packaged meat product in order to determine the appropriate preservation method.
Macián
M C
MC
Departamento de Microbiología y Ecología, Universitat de València, Spain.
Chenoll
E
E
Aznar
R
R
D000818:Animals; D001483:Base Sequence; D002645:Chickens; D017931:DNA Primers / Q000032:analysis; D004269:DNA, Bacterial / Q000032:analysis; D007777:Lactobacillaceae / Q000302:isolation & purification*; D007934:Leuconostoc / Q000302:isolation & purification*; D008461:Meat Products / Q000382:microbiology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D016415:Sequence Alignment; D013552:Swine; D014422:Turkeys
D017931:DNA Primers; D004269:DNA, Bacterial