Microbiota of deciduous endodontic infections analysed by MDA and Checkerboard DNA-DNA hybridization

Int Endod J. 2011 Mar;44(3):225-35. doi: 10.1111/j.1365-2591.2010.01805.x. Epub 2010 Nov 17.

Abstract

Aims: To evaluate the microbiota of endodontic infections in deciduous teeth by Checkerboard DNA-DNA hybridization after uniform amplification of DNA in samples by multiple displacement amplification (MDA).

Methodology: Forty samples from the root canal system of deciduous teeth exhibiting pulp necrosis with or without radiographically detectable periradicular/interradicular bone resorption were collected and 32 were analysed, with three individuals contributing two samples; these were MDA-amplified and analysed by Checkerboard DNA-DNA hybridization for levels of 83 bacterial taxa. Two outcome measures were used: the percentage of teeth colonized by each species and the mean proportion of each bacterial taxon present across all samples.

Results: The mean amount of DNA in the samples prior to amplification was 5.2 (±4.7) ng and 6.1 (±2.3) μg after MDA. The mean number of species detected per sample was 19 (±4) (range: 3-66) to the nearest whole number. The most prevalent taxa were Prevotella intermedia (96.9%), Neisseria mucosa (65.6%), Prevotella nigrescens (56.2%) and Tannerella forsythia (56.2%). Aggregatibacter (Haemophilus) aphrophilus and Helicobacter pylori were not detected. P. intermedia (10%), Prevotella tannerae (7%) and Prevotella nigrescens (4.3%) presented the highest mean proportions of the target species averaged across the positive samples.

Conclusion: Root canals of infected deciduous teeth had a diverse bacterial population. Prevotella sp. were commonly found with P. intermedia, Prevotella tannerae and Prevotella nigrescens amongst the most prominent species detected.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacteria / classification*
  • Bacteria / genetics
  • Bacteria / isolation & purification
  • Bacteriological Techniques / methods
  • Child
  • Child, Preschool
  • Colony Count, Microbial
  • DNA, Bacterial / analysis*
  • Dental Pulp Cavity / microbiology*
  • Dental Pulp Necrosis / microbiology*
  • Humans
  • Nucleic Acid Amplification Techniques / methods
  • Nucleic Acid Hybridization / methods
  • Tooth, Deciduous / microbiology*

Substances

  • DNA, Bacterial