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PMID: 10618239 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Detection and identification of gastrointestinal Lactobacillus species by using denaturing gradient gel electrophoresis and species-specific PCR primers.

Applied and environmental microbiology ·Vol. 66 ·No. 1 ·2000-01-00 ·Pages 297-303

Walter J, Tannock GW, Tilsala-Timisjarvi A, Rodtong S, Loach DM, Munro K, Alatossava T

Abstract

Denaturing gradient gel electrophoresis (DGGE) of DNA fragments obtained by PCR amplification of the V2-V3 region of the 16S rRNA gene was used to detect the presence of Lactobacillus species in the stomach contents of mice. Lactobacillus isolates cultured from human and porcine gastrointestinal samples were identified to the species level by using a combination of DGGE and species-specific PCR primers that targeted 16S-23S rRNA intergenic spacer region or 16S rRNA gene sequences. The identifications obtained by this approach were confirmed by sequencing the V2-V3 region of the 16S rRNA gene and by a BLAST search of the GenBank database.

MeSH Terms
Animals DNA Primers DNA, Bacterial/analysis DNA, Ribosomal/analysis Electrophoresis, Polyacrylamide Gel/methods Genes, rRNA Humans Lactobacillus/classification,genetics,isolation & purification Mice Polymerase Chain Reaction/methods RNA, Ribosomal, 16S/genetics RNA, Ribosomal, 23S/genetics Species Specificity Stomach/microbiology
Chemicals
DNA Primers DNA, Bacterial DNA, Ribosomal RNA, Ribosomal, 16S RNA, Ribosomal, 23S
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Walter J
Department of Microbiology, University of Otago, Dunedin, New Zealand.
Tannock G W
Tilsala-Timisjarvi A
Rodtong S
Loach D M
Munro K
Alatossava T
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10 references, click to expand
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Article Info
Journal
Applied and environmental microbiology
Abbr.
Appl Environ Microbiol
ISSN
0099-2240
Published
2000-01-00
Pages
297-303
Language
English
Region
United States
NLM ID
7605801
PMCID
PMC91821
Subset
IM
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