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Applied and Environmental Microbiology, January 2004, p. 167-173, Vol. 70, No. 1
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.1.167-173.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Quantitative PCR with 16S rRNA-Gene-Targeted Species-Specific Primers for Analysis of Human Intestinal Bifidobacteria

Takahiro Matsuki,* Koichi Watanabe, Junji Fujimoto, Yukiko Kado, Toshihiko Takada, Kazumasa Matsumoto, and Ryuichiro Tanaka

Yakult Central Institute for Microbiological Research, Kunitachi, Tokyo 186-8650, Japan

Received 18 June 2003/ Accepted 6 October 2003

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 106 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 >106 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.

* Corresponding author. Mailing address: Yakult Central Institute for Microbiological Research, 1796 Yaho, Kunitachi, Tokyo 186-8650, Japan. Phone: 81 (42) 577 8962. Fax: 81 (42) 577 3020. E-mail: takahiro-matsuki{at}yakult.co.jp.

Applied and Environmental Microbiology, January 2004, p. 167-173, Vol. 70, No. 1
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.1.167-173.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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