Identification, detection, and enumeration of human bifidobacterium species by PCR targeting the transaldolase gene.
Methods that enabled the identification, detection, and enumeration of Bifidobacterium species by PCR targeting the transaldolase gene were tested. Bifidobacterial species isolated from the feces of human adults and babies were identified by PCR amplification of a 301-bp transaldolase gene sequence and comparison of the relative migrations of the DNA fragments in denaturing gradient gel electrophoresis (DGGE). Two subtypes of Bifidobacterium longum, five subtypes of Bifidobacterium adolescentis, and two subtypes of Bifidobacterium pseudocatenulatum could be differentiated using PCR-DGGE. Bifidobacterium angulatum and B. catenulatum type cultures could not be differentiated from each other. Bifidobacterial species were also detected directly in fecal samples by this combination of PCR and DGGE. The number of species detected was less than that detected by PCR using species-specific primers targeting 16S ribosomal DNA (rDNA). Real-time quantitative PCR targeting a 110-bp transaldolase gene sequence was used to enumerate bifidobacteria in fecal samples. Real-time quantitative PCR measurements of bifidobacteria in fecal samples from adults correlated well with results obtained by culture when either a 16S rDNA sequence or the transaldolase gene sequence was targeted. In the case of samples from infants, 16S rDNA-targeted PCR was superior to PCR targeting the transaldolase gene for the quantification of bifidobacterial populations.
Requena
Teresa
T
Department of Microbiology, University of Otago, Dunedin, New Zealand.
Burton
Jeremy
J
Matsuki
Takahiro
T
Munro
Karen
K
Simon
Mary Alice
MA
Tanaka
Ryuichiro
R
Watanabe
Koichi
K
Tannock
Gerald W
GW
D001483:Base Sequence; D001644:Bifidobacterium / Q000235:genetics / Q000302:isolation & purification*; D003199:Computer Systems; D004269:DNA, Bacterial / Q000032:analysis; D005243:Feces / Q000382:microbiology; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012689:Sequence Homology, Nucleic Acid; D014153:Transaldolase / Q000235:genetics*
D004269:DNA, Bacterial; D014153:Transaldolase