title: Quantitative PCR with 16S rRNA-gene-targeted species-specific primers for analysis of human intestinal bifidobacteria.
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.
authors.affiliation: Yakult Central Institute for Microbiological Research, Kunitachi, Tokyo 186-8650, Japan.
authors.forename: Takahiro
authors.initials: T
authors.lastname: Matsuki
authors.forename: Koichi
authors.initials: K
authors.lastname: Watanabe
authors.forename: Junji
authors.initials: J
authors.lastname: Fujimoto
authors.forename: Yukiko
authors.initials: Y
authors.lastname: Kado
authors.forename: Toshihiko
authors.initials: T
authors.lastname: Takada
authors.forename: Kazumasa
authors.initials: K
authors.lastname: Matsumoto
authors.forename: Ryuichiro
authors.initials: R
authors.lastname: Tanaka
mesh_terms: D000328:Adult; D001483:Base Sequence; D001644:Bifidobacterium / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D015169:Colony Count, Microbial; D017931:DNA Primers; D004275:DNA, Ribosomal / Q000032:analysis; D005260:Female; D020459:Genes, rRNA; D006801:Humans; D007422:Intestines / Q000382:microbiology*; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012336:RNA, Ribosomal, 16S / Q000235:genetics*; D017422:Sequence Analysis, DNA; D013045:Species Specificity
chemical_list: D017931:DNA Primers; D004275:DNA, Ribosomal; D012336:RNA, Ribosomal, 16S