Detection and identification of gastrointestinal Lactobacillus species by using denaturing gradient gel electrophoresis and species-specific PCR primers.
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.
Walter
J
J
Department of Microbiology, University of Otago, Dunedin, New Zealand.
Tannock
G W
GW
Tilsala-Timisjarvi
A
A
Rodtong
S
S
Loach
D M
DM
Munro
K
K
Alatossava
T
T
D000818:Animals; D017931:DNA Primers; D004269:DNA, Bacterial / Q000032:analysis; D004275:DNA, Ribosomal / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D020459:Genes, rRNA; D006801:Humans; D007778:Lactobacillus / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D051379:Mice; D016133:Polymerase Chain Reaction / Q000379:methods; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012338:RNA, Ribosomal, 23S / Q000235:genetics; D013045:Species Specificity; D013270:Stomach / Q000382:microbiology*
D017931:DNA Primers; D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D012336:RNA, Ribosomal, 16S; D012338:RNA, Ribosomal, 23S