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Applied and Environmental Microbiology, April 2009, p. 2573-2576, Vol. 75, No. 8
0099-2240/09/$08.00+0     doi:10.1128/AEM.02576-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Hierarchical Oligonucleotide Primer Extension as a Time- and Cost-Effective Approach for Quantitative Determination of Bifidobacterium spp. in Infant Feces{triangledown}

Pei-Ying Hong,1 Gaik Chin Yap,2 Bee Wah Lee,2 Kaw Yan Chua,2 and Wen-Tso Liu1,3*

Division of Environmental Science and Engineering, National University of Singapore, Singapore 117576,1 Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117576,2 Department of Civil and Environmental Engineering, University of Illinois at Urbana-Champaign, Urbana, Illinois 618013

Received 11 November 2008/ Accepted 2 February 2009

The Bifidobacterium spp. present in 10 infant fecal samples (4 from infants with eczema and 6 from healthy infants) were quantified with both hierarchical oligonucleotide primer extension (HOPE) and fluorescence in situ hybridization-flow cytometry. The relative abundances of Bifidobacterium longum and B. catenulatum with respect to the total bifidobacteria had a poor correlation ({rho}, <0.600; P value, >0.208), presumably due to differences in primer specificity and the level of hybridization stringency of both methods. In contrast, the relative abundances of organisms of the genus Bifidobacterium against the total amplified 16S rRNA genes and those of B. adolescentis, B. bifidum, and B. breve against the genus Bifidobacterium exhibited a good statistical correlation ({rho}, >0.783; P value, <0.066). This good comparability supports HOPE as a method to achieve high-throughput quantitative determination of bacterial targets in a time- and cost-effective manner.

* Corresponding author. Mailing address: Department of Civil & Environmental Engineering, University of Illinois at Urbana-Champaign, 3207 Newmark Civil Engineering Laboratory, 205 North Mathews Ave., Urbana, IL 61801-2352. Phone: (217) 333-8442. Fax: (217) 333-6968. E-mail: wtliu{at}illinois.edu

{triangledown} Published ahead of print on 13 February 2009.

Applied and Environmental Microbiology, April 2009, p. 2573-2576, Vol. 75, No. 8
0099-2240/09/$08.00+0     doi:10.1128/AEM.02576-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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