A TaqMan-based real-time PCR assay for the specific detection and quantification of Leuconostoc mesenteroides in meat products.
A new real-time PCR procedure was developed for the specific detection and quantification of Leuconostoc mesenteroides in meat products. It is a TaqMan assay based on 23S rRNA gene targeted primers and probe. Specificity was evaluated using purified DNA from 132 strains: 102 lactic acid bacteria (LAB), including 57 reference strains and 46 food isolates, belonging to genus Leuconostoc and related genera, and 30 non-LAB strains. Quantification was linear over at least 5 log units using both serial dilutions of purified DNA and calibrated cell suspensions from Leuconostoc mesenteroides ssp. dextranicum CECT 912T. This assay was able to detect at least five genomic equivalents, using purified DNA or 59 CFU per reaction when using calibrated cell suspensions. It performed successfully when tested on an artificially inoculated meat product, with a minimum threshold of 10(4) CFU g(-1) for the accurate quantification of Leuconostoc mesenteroides.
Elizaquível
Patricia
P
Departamento de Microbiología, Universitat de València, Burjassot, Valencia, Spain.
Chenoll
Empar
E
Aznar
Rosa
R
D000818:Animals; D017931:DNA Primers; D004269:DNA, Bacterial / Q000235:genetics; D007777:Lactobacillaceae / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification*; D008461:Meat Products / Q000382:microbiology*; D008969:Molecular Sequence Data; D012338:RNA, Ribosomal, 23S / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA
D017931:DNA Primers; D004269:DNA, Bacterial; D012338:RNA, Ribosomal, 23S