Polymerase chain reaction-based denaturing gradient gel electrophoresis in the evaluationof oral microbiota

Y. Li, D. Saxena, V. M. Barnes, H. M. Trivedi, Y. Ge, T. Xu

Research output: Contribution to journalArticle

Abstract

Background/aims: Clinical evaluation of oral microbial reduction after a standard prophylactic treatment has traditionally been based on bacterial cultivation methods. However, not all microbes in saliva or dental plaque can be cultivated. Polymerase chain reaction-based denaturing gradient gel electrophoresis (PCR-DGGE) is a cultivation-independent molecular fingerprinting technique that allows the assessment of the predominant bacterial species present in the oral cavity. This study sought to evaluate the oral microbial changes that occurred after a standard prophylactic treatment with a conventional oral care product using PCR-DGGE. Methods: Twelve healthy adults participated in the study. Pooled plaque samples were collected at baseline, 24 h after prophylaxis (T1), and 4 days after toothbrushing with fluoride toothpaste (T4). The total microbial genomic DNA of the plaque was isolated. PCR was performed with a set of universal bacterial 16S rDNA primers. The PCR-amplified 16S rDNA fragments were separated by DGGE. The effects of the treatment and of dental brushing were assessed by comparing the PCR-DGGE fingerprinting profiles. Results: The mean numbers of detected PCR amplicons were 22.3 ± 6.1 for the baseline group, 13.0 ± 3.1 for the T1 group, and 13.5 ± 4.3 for the T4 group; the differences among the three groups were statistically significant (P < 0.01). The study also found a significant difference in the mean similarities of microbial profiles between the baseline and the treatment groups (P < 0.001). Conclusion: PCR-based DGGE has been shown to be an excellent means of rapidly and accurately assessing oral microbial changes in this clinical study.

Original languageEnglish (US)
Pages (from-to)333-339
Number of pages7
JournalOral Microbiology and Immunology
Volume21
Issue number5
DOIs
StatePublished - Oct 2006

Keywords

  • Dental plaque
  • Fluoride dentifrice
  • Oral microbes
  • Polymerase chain reaction-denaturing gradient gel electrophoresis
  • Prophylactic treatment

ASJC Scopus subject areas

  • Microbiology
  • Immunology
  • Dentistry(all)
  • Microbiology (medical)

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