Use of the genomic subtractive hybridization technique to develop a real-time PCR assay for quantitative detection of Prevotella spp. in oral biofilm samples

J Clin Microbiol. 2005 Jun;43(6):2948-51. doi: 10.1128/JCM.43.6.2948-2951.2005.

Abstract

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.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacteroidaceae Infections / microbiology
  • Biofilms / growth & development*
  • DNA Probes
  • DNA, Bacterial / analysis
  • Dental Plaque / microbiology
  • Humans
  • Mouth / microbiology*
  • Nucleic Acid Hybridization / methods*
  • Polymerase Chain Reaction / methods*
  • Prevotella / classification
  • Prevotella / genetics
  • Prevotella / isolation & purification*
  • Species Specificity
  • Taq Polymerase

Substances

  • DNA Probes
  • DNA, Bacterial
  • Taq Polymerase