Affiliation:
1. Department of Medical Microbiology, University of Groningen, 9700 RB Groningen, The Netherlands
Abstract
ABSTRACT
Six 16S rRNA-targeted oligonucleotide probes were designed, validated, and used to quantify predominant groups of anaerobic bacteria in human fecal samples. A set of two probes was specific for species of the
Bacteroides fragilis
group and the species
Bacteroides distasonis
. Two others were designed to detect species of the
Clostridium histolyticum
and the
Clostridium lituseburense
groups. Another probe was designed for the genera
Streptococcus
and
Lactococcus
, and the final probe was designed for the species of the
Clostridium coccoides-Eubacterium rectale
group. The temperature of dissociation of each of the probes was determined. The specificities of the probes for a collection of target and reference organisms were tested by dot blot hybridization and fluorescent in situ hybridization (FISH). The new probes were used in initial FISH experiments to enumerate human fecal bacteria. The combination of the two
Bacteroides
-specific probes detected a mean of 5.4 × 10
10
cells per g (dry weight) of feces; the
Clostridium coccoides-Eubacterium rectale
group-specific probe detected a mean of 7.2 × 10
10
cells per g (dry weight) of feces. The
Clostridium histolyticum
,
Clostridium lituseburense
, and
Streptococcus-Lactococcus
group-specific probes detected only numbers of cells ranging from 1 × 10
7
to 7 × 10
8
per g (dry weight) of feces. Three of the newly designed probes and three additional probes were used in further FISH experiments to study the fecal flora composition of nine volunteers over a period of 8 months. The combination of probes was able to detect at least two-thirds of the fecal flora. The normal biological variations within the fecal populations of the volunteers were determined and indicated that these variations should be considered when evaluating the effects of agents modulating the flora.
Publisher
American Society for Microbiology
Subject
Ecology,Applied Microbiology and Biotechnology,Food Science,Biotechnology
Cited by
822 articles.
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