Back to Search
Start Over
Development and use of tuf gene-based primers for the multiplex PCR detection of Lactobacillus acidophilus, Lactobacillus casei group, Lactobacillus delbrueckii, and Bifidobacterium longum in commercial dairy products
- Source :
- Journal of food protection. 72(1)
- Publication Year :
- 2009
-
Abstract
- PCR primers specific for the detection of Lactobacillus acidophilus, Lactobacillus casei group, Lactobacillus delbrueckii, and Bifidobacterium longum were designed based on the elongation factor Tu gene (tuf). The specificity of these four primer sets were confirmed by PCR with 88 bacterial strains of Lactobacillus, Enterococcus, Bifidobacterium, and other bacterial species. Results indicated that these primer sets generated predicted PCR products of 397, 230, 202, and 161 bp for L. acidophilus, L. delbrueckii, L. casei group, and B. longum, respectively. Bacterial species other than the target organisms tested did not generate false-positive results. When these four primer sets were combined for the simultaneous detection of the lactic acid bacteria (LAB) in fermented milk products including yogurt, the LAB species listed on the labels of these products could be identified without the preenrichment step. The identification limit for each LAB strain with this multiplex PCR method was N X 10(3) CFU/ml in milk samples. The results of our multiplex PCR method were confirmed by PCR assay using primers based on the 16S rDNA or the 16S-23S intergenic spacer region and by biochemical tests using the API 50 CHL kit. When this multiplex PCR method was used with the determination of counts of total viable LAB and bifidobacteria, the quality of commercial fermented milk products could be assured.
- Subjects :
- DNA, Bacterial
Lactobacillus casei
Bifidobacterium longum
Time Factors
Cultured Milk Products
Colony Count, Microbial
Microbiology
Polymerase Chain Reaction
Sensitivity and Specificity
fluids and secretions
Lactobacillus acidophilus
Species Specificity
Lactobacillus
RNA, Ribosomal, 16S
Multiplex polymerase chain reaction
DNA, Ribosomal Spacer
Fermented milk products
Animals
Humans
Bifidobacterium
DNA Primers
Lactobacillus delbrueckii
biology
Probiotics
food and beverages
Sequence Analysis, DNA
biology.organism_classification
Bacterial Typing Techniques
Molecular Weight
Lacticaseibacillus casei
RNA, Ribosomal, 23S
Enterococcus
Dairy Products
Food Science
Subjects
Details
- ISSN :
- 0362028X
- Volume :
- 72
- Issue :
- 1
- Database :
- OpenAIRE
- Journal :
- Journal of food protection
- Accession number :
- edsair.doi.dedup.....e8fc68a9b9fa221204b5bd858151b71b