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Quantitative PCR with 16S rRNA-gene-targeted species-specific primers for analysis of human intestinal bifidobacteria.
- Source :
-
Applied and environmental microbiology [Appl Environ Microbiol] 2004 Jan; Vol. 70 (1), pp. 167-73. - Publication Year :
- 2004
-
Abstract
- A highly sensitive quantitative PCR detection method has been developed and applied to the distribution analysis of human intestinal bifidobacteria by combining real-time PCR with Bifidobacterium genus- and species-specific primers. Real-time PCR detection of serially diluted DNA extracted from cultured bifidobacteria was linear for cell counts ranging from 10(6) to 10 cells per PCR assay. It was also found that the method was applicable to the detection of Bifidobacterium in feces when it was present at concentrations of >10(6) cells per g of feces. Concerning the distribution of Bifidobacterium species in intestinal flora, the Bifidobacterium adolescentis group, the Bifidobacterium catenulatum group, and Bifidobacterium longum were found to be the three predominant species by examination of DNA extracted from the feces of 46 healthy adults. We also examined changes in the population and composition of Bifidobacterium species in human intestinal flora of six healthy adults over an 8-month period. The results showed that the composition of bifidobacterial flora was basically stable throughout the test period.
- Subjects :
- Adult
Base Sequence
Bifidobacterium genetics
Colony Count, Microbial
DNA, Ribosomal analysis
Female
Genes, rRNA
Humans
Male
Middle Aged
Molecular Sequence Data
Sequence Analysis, DNA
Species Specificity
Bifidobacterium classification
Bifidobacterium isolation & purification
DNA Primers
Intestines microbiology
Polymerase Chain Reaction methods
RNA, Ribosomal, 16S genetics
Subjects
Details
- Language :
- English
- ISSN :
- 0099-2240
- Volume :
- 70
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- Applied and environmental microbiology
- Publication Type :
- Academic Journal
- Accession number :
- 14711639
- Full Text :
- https://doi.org/10.1128/AEM.70.1.167-173.2004