Checkerboard DNA-DNA hybridisation technology focused on the analysis of Gram-positive cariogenic bacteria

J Microbiol Methods. 2002 Nov;51(3):301-11. doi: 10.1016/s0167-7012(02)00106-9.

Abstract

Checkerboard DNA-DNA hybridisation enabled the quantitative analysis of plaque samples against 40 microbial species simultaneously, using digoxygenin-labelled, whole-genome DNA probes. This technique was initially developed to study the predominantly Gram-negative sub-gingival plaque microbiota. The aim of this study was to apply it to a suite of predominantly Gram-positive microorganisms, such as those implicated in cariogenesis. To specifically target Gram-positive species (and Candida albicans) required optimisation and modification of DNA extraction, prehybridisation, hybridisation, and antibody detection conditions. The suitability of the revised technique for clinical and epidemiological studies was confirmed using interproximal plaque from small groups of 5- to 6-year-old children of high (decayed, missing, or filled teeth (dmft)> or =5, n=8) and zero (n=5) caries rates.

Publication types

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

MeSH terms

  • Buffers
  • Candida albicans / classification
  • Candida albicans / genetics
  • Child
  • Child, Preschool
  • DNA Probes
  • DNA, Bacterial / analysis*
  • DNA, Bacterial / isolation & purification
  • DNA, Fungal / analysis
  • DNA, Fungal / isolation & purification
  • Dental Caries / microbiology*
  • Dental Plaque / microbiology*
  • Digoxigenin
  • Gram-Positive Bacteria / classification
  • Gram-Positive Bacteria / genetics
  • Humans
  • Nucleic Acid Hybridization / methods*
  • Sensitivity and Specificity

Substances

  • Buffers
  • DNA Probes
  • DNA, Bacterial
  • DNA, Fungal
  • Digoxigenin