Affiliation:
1. Laboratory of Microbiology, Wageningen University, 6703 CT Wageningen
2. Wageningen Center for Food Sciences, 6700 AN Wageningen, The Netherlands
3. Department of Gastroenterology, HEGP, 75908 Paris Cedex 50, France
Abstract
ABSTRACT
A
Lactobacillus
group-specific PCR primer, S-G-Lab-0677-a-A-17, was developed to selectively amplify 16S ribosomal DNA (rDNA) from lactobacilli and related lactic acid bacteria, including members of the genera
Leuconostoc
,
Pediococcus
, and
Weissella.
Amplicons generated by PCR from a variety of gastrointestinal (GI) tract samples, including those originating from feces and cecum, resulted predominantly in
Lactobacillus
-like sequences, of which ca. 28% were most similar to the 16S rDNA of
Lactobacillus ruminis
. Moreover, four sequences of
Leuconostoc
species were retrieved that, so far, have only been detected in environments other than the GI tract, such as fermented food products. The validity of the primer was further demonstrated by using
Lactobacillus
-specific PCR and denaturing gradient gel electrophoresis (DGGE) of the 16S rDNA amplicons of fecal and cecal origin from different age groups. The stability of the GI-tract bacterial community in different age groups over various time periods was studied. The
Lactobacillus
community in three adults over a 2-year period showed variation in composition and stability depending on the individual, while successional change of the
Lactobacillus
community was observed during the first 5 months of an infant’s life. Furthermore, the specific PCR and DGGE approach was tested to study the retention in fecal samples of a
Lactobacillus
strain administered during a clinical trial. In conclusion, the combination of specific PCR and DGGE analysis of 16S rDNA amplicons allows the diversity of important groups of bacteria that are present in low numbers in specific ecosystems to be characterized, such as the lactobacilli in the human GI tract.
Publisher
American Society for Microbiology
Subject
Ecology,Applied Microbiology and Biotechnology,Food Science,Biotechnology
Cited by
572 articles.
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