DESIGN AND EVALUATION OF A 16S RIBOSOMAL-RNA-TARGETED OLIGONUCLEOTIDEPROBE FOR SPECIFIC DETECTION AND QUANTITATION OF HUMAN FECAL BACTEROIDES POPULATIONS
J. Dore et al., DESIGN AND EVALUATION OF A 16S RIBOSOMAL-RNA-TARGETED OLIGONUCLEOTIDEPROBE FOR SPECIFIC DETECTION AND QUANTITATION OF HUMAN FECAL BACTEROIDES POPULATIONS, Systematic and applied microbiology, 21(1), 1998, pp. 65-71
Colonic Bacteroides include several species which, by their population
level and activities, are significant contributers to the metabolic a
ctivity and health of man and animals. Yet, the understanding of their
ecology has been hampered by the lack of highly specific and reliable
enumeration techniques. Based on 16S rRNA sequence comparisons within
the available database, we have designed an 18-mer oligonucleotide th
at targets a region common to and specific for the Bacteroides-Porphyr
omonas-Prevotella group. We have tested the specificity of the probe a
nd its usefulness for studies of human faecal samples. Under experimen
tally optimized hybridization conditions, the probe was shown to simil
arly recognize the rDNA obtained from 40 strains representing 8 specie
s of the Bacteroides-Porphyromonas-Prevotella group. Importantly, it d
id not recognize 31 strains of microorganisms representing 8 genera of
the dominant human faecal microbiota. Among selected colonies of domi
nant microorganisms of the faecal flora of two human individuals, stra
ins identified as B. vulgatus by immunoblots using a species-specific
monoclonal antibody were ail detected by the probe. Colony hybridizati
on was used to enumerate total Bacteroides-group microorganisms in fae
cal specimen from children and adults. The probe described therein was
further used in quantitative RNA blots to monitor fluctuations of the
Bacteroides-group versus Bifidobacterium genus in frozen faecal sampl
es from a child between 85 and 125 days of age. It will be applicable
to similar investigations of other anaerobic environments.