Identification of oral species of the genus Veillonella by polymerase chain reaction.
BACKGROUND
Members of the genus Veillonella cannot be reliably distinguished by their biochemical characteristics and phenotypic features. Moreover, DNA-DNA hybridization and sequence analyses of the 16S ribosomal RNA gene including random fragment length polymorphism analysis, are complex and time-consuming procedures that are not well-suited to identifying oral species of Veillonella: Veillonella atypica, Veillonella denticariosi, Veillonella dispar, Veillonella parvula, and Veillonella rogosae.


METHODS
In this study, five forward primers and a reverse primer were designed for polymerase chain reaction (PCR) according to the partial sequences of the rpoB genes of these oral Veillonella species.


RESULTS
The forward primers were species-specific for these five Veillonella species, and could produce specific amplicons when used together with reverse primer and individual DNA templates of these species in PCR. These primer pairs were also found to discriminate between the respective species, and the Veillonella strains isolated from human oral cavities were successfully assigned to one of the five oral species of the genus Veillonella based on their specific products by PCR.


CONCLUSIONS
A simple two-step PCR procedure using the five sets of primer pairs developed in the present study is a rapid and reliable method for the identification of the recognized oral Veillonella species.
Igarashi
E
E
Department of Oral Microbiology, School of Dentistry, Health Sciences University of Hokkaido, Ishikari-Tobetsu, Hokkaido 061-0293, Japan.
Kamaguchi
A
A
Fujita
M
M
Miyakawa
H
H
Nakazawa
F
F
D001426:Bacterial Proteins; D015373:Bacterial Typing Techniques; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000032:analysis; D005798:Genes, Bacterial; D006801:Humans; D009055:Mouth / Q000382:microbiology*; D016133:Polymerase Chain Reaction / Q000379:methods; D017422:Sequence Analysis, DNA / Q000379:methods; D013045:Species Specificity; D014678:Veillonella / Q000145:classification / Q000302:isolation & purification*
D001426:Bacterial Proteins; D017931:DNA Primers; D004269:DNA, Bacterial