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Applied and Environmental Microbiology, May 2002, p. 2420-2427, Vol. 68, No. 5
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.5.2420-2427.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Identification, Detection, and Enumeration of Human Bifidobacterium Species by PCR Targeting the Transaldolase Gene

Teresa Requena,1,{dagger} Jeremy Burton,1 Takahiro Matsuki,2 Karen Munro,1 Mary Alice Simon,1 Ryuichiro Tanaka,2 Koichi Watanabe,2 and Gerald W. Tannock1*

Department of Microbiology, University of Otago, Dunedin, New Zealand,1 Yakult Central Institute for Microbiological Research, Tokyo, Japan2

Received 24 September 2001/ Accepted 14 January 2002

Methods that enabled the identification, detection, and enumeration of Bifidobacterium species by PCR targeting the transaldolase gene were tested. Bifidobacterial species isolated from the feces of human adults and babies were identified by PCR amplification of a 301-bp transaldolase gene sequence and comparison of the relative migrations of the DNA fragments in denaturing gradient gel electrophoresis (DGGE). Two subtypes of Bifidobacterium longum, five subtypes of Bifidobacterium adolescentis, and two subtypes of Bifidobacterium pseudocatenulatum could be differentiated using PCR-DGGE. Bifidobacterium angulatum and B. catenulatum type cultures could not be differentiated from each other. Bifidobacterial species were also detected directly in fecal samples by this combination of PCR and DGGE. The number of species detected was less than that detected by PCR using species-specific primers targeting 16S ribosomal DNA (rDNA). Real-time quantitative PCR targeting a 110-bp transaldolase gene sequence was used to enumerate bifidobacteria in fecal samples. Real-time quantitative PCR measurements of bifidobacteria in fecal samples from adults correlated well with results obtained by culture when either a 16S rDNA sequence or the transaldolase gene sequence was targeted. In the case of samples from infants, 16S rDNA-targeted PCR was superior to PCR targeting the transaldolase gene for the quantification of bifidobacterial populations.

* Corresponding author. Mailing address: Department of Microbiology, University of Otago, P.O. Box 56, Dunedin, New Zealand. Phone: 64-3-479-7713. Fax: 64-3-479-8540. E-mail: gerald.tannock{at}stonebow.otago.ac.nz.

{dagger} Present address: Instituto del Frio (CSIC), Ciudad Universitaria, 28040 Madrid, Spain.

Applied and Environmental Microbiology, May 2002, p. 2420-2427, Vol. 68, No. 5
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.5.2420-2427.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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