Use of the genomic subtractive hybridization technique to develop a real-time PCR assay for quantitative detection of Prevotella spp. in oral biofilm samples.
Genomic subtractive hybridization was used to design Prevotella nigrescens-specific primers and TaqMan probes. Based on this technique, a TaqMan real-time PCR assay was developed for quantifying four oral black-pigmented Prevotella species. The combination of real-time PCR and genomic subtractive hybridization is useful for preparing species-specific primer-probe sets for closely related species.
Nagashima
Shiori
S
Division of Community Oral Health Science, Kyushu Dental College, 2-6-1 Manazuru, Kitakyushu 803-8580, Japan.
Yoshida
Akihiro
A
Suzuki
Nao
N
Ansai
Toshihiro
T
Takehara
Tadamichi
T
D016866:Bacteroidaceae Infections / Q000382:microbiology; D018441:Biofilms / Q000254:growth & development*; D015342:DNA Probes; D004269:DNA, Bacterial / Q000032:analysis; D003773:Dental Plaque / Q000382:microbiology; D006801:Humans; D009055:Mouth / Q000382:microbiology*; D009693:Nucleic Acid Hybridization / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D018720:Prevotella / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D013045:Species Specificity; D019914:Taq Polymerase
D015342:DNA Probes; D004269:DNA, Bacterial; D019914:Taq Polymerase