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Applied and Environmental Microbiology, January 2002, p. 114-123, Vol. 68, No. 1
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.1.114-123.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Molecular Diversity of Lactobacillus spp. and Other Lactic Acid Bacteria in the Human Intestine as Determined by Specific Amplification of 16S Ribosomal DNA

Hans G.H.J. Heilig,1 Erwin G. Zoetendal,1,2 Elaine E. Vaughan,1,2* Philippe Marteau,3 Antoon D.L. Akkermans,1 and Willem M. de Vos1,2

Laboratory of Microbiology, Wageningen University, 6703 CT Wageningen,1 Wageningen Center for Food Sciences, 6700 AN Wageningen, The Netherlands,2 Department of Gastroenterology, HEGP, 75908 Paris Cedex 50, France3

Received 24 April 2001/ Accepted 20 September 2001

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.

* Corresponding author. Mailing address: Laboratory of Microbiology, Wageningen University, Hesselink van Suchtelenweg 4, 6703 CT Wageningen, The Netherlands. Phone: 31-317-483113. Fax: 31-317-483829. Email: elaine.vaughan{at}algemeen.micr.wau.nl.

Applied and Environmental Microbiology, January 2002, p. 114-123, Vol. 68, No. 1
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.1.114-123.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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