Open AccessHierarchical Oligonucleotide Primer Extension as a Time- and Cost-Effective Approach for Quantitative Determination of Bifidobacterium spp. in Infant Feces
Author(s) -
PeiYing Hong,
Gaik Chin Yap,
Bee Wah Lee,
Kaw Yan Chua,
Wen Tso Liu
Publication year - 2009
Publication title -
applied and environmental microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.552
H-Index - 324
eISSN - 1070-6291
pISSN - 0099-2240
DOI - 10.1128/aem.02576-08
Subject(s) - biology , bifidobacterium , bifidobacterium bifidum , primer extension , bifidobacterium breve , primer (cosmetics) , actinomycetaceae , oligonucleotide , 16s ribosomal rna , feces , microbiology and biotechnology , genetics , dna , bacteria , lactobacillus , chemistry , base sequence , organic chemistry
TheBifidobacterium 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 ofBifidobacterium longum andB .catenulatum with respect to the total bifidobacteria had a poor correlation (ρ, <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 genusBifidobacterium against the total amplified 16S rRNA genes and those ofB .adolescentis ,B .bifidum , andB .breve against the genusBifidobacterium exhibited a good statistical correlation (ρ, >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.