Application of denaturing gradient gel electrophoresis (DGGE) to the analysis of microbial communities of subgingival plaque

C. Fujimoto, H. Maeda, S. Kokeguchi, S. Takashiba, Fusanori Nishimura, H. Arai, K. Fukui, Y. Murayama

Research output: Contribution to journalArticle

51 Citations (Scopus)

Abstract

Objectives: Denaturing gradient gel electrophoresis (DGGE) was applied to the microbiologic examination of subgingival plaque. Materials and methods: The PCR primers were designed from conserved nucleotide sequences on 16S ribosomal RNA gene (16SrDNA) with GC rich clamp at the 5′-end. Polymerase chain reaction (PCR) was performed using the primers and genomic DNAs of typical periodontal bacteria. The generated 16SrDNA fragments were separated by denaturing gel. Results: Although the sizes of the amplified DNA fragments were almost the same among the species, 16SrDNAs of the periodontal bacteria were distinguished according to their specific sequences. The microflora of clinical plaque samples were profiled by the PCR-DGGE method, and the dominant 16SrDNA bands were cloned and sequenced. Simultaneously, Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis and Prevotella intermedia were detected by an ordinary PCR method. In the deep periodontal pockets, the bacterial community structures were complicated and P. gingivalis was the most dominant species, whereas the DGGE profiles were simple and Streptococcus or Neisseria species were dominant in the shallow pockets. The species-specific PCR method revealed the presence of A. actinomycetemcomitans, P. gingivalis and P. intermedia in the clinical samples. However, corresponding bands were not always observed in the DGGE profiles, indicating a lower sensitivity of the DGGE method. Conclusion: Although the DGGE method may have a lower sensitivity than the ordinary PCR methods, it could visualize the bacterial qualitative compositions and reveal the major species of the plaque. The DGGE analysis and following sequencing may have the potential to be a promising bacterial examination procedure in periodontal diseases.

Original languageEnglish
Pages (from-to)440-445
Number of pages6
JournalJournal of Periodontal Research
Volume38
Issue number4
DOIs
Publication statusPublished - Jan 1 2003
Externally publishedYes

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Denaturing Gradient Gel Electrophoresis
Polymerase Chain Reaction
Porphyromonas gingivalis
Prevotella intermedia
Aggregatibacter actinomycetemcomitans
16S Ribosomal RNA
Periodontal Pocket
Bacteria
Neisseria
Bacterial Structures
DNA Primers
Conserved Sequence
Periodontal Diseases
Streptococcus
rRNA Genes
Gels
DNA

All Science Journal Classification (ASJC) codes

  • Periodontics

Cite this

Application of denaturing gradient gel electrophoresis (DGGE) to the analysis of microbial communities of subgingival plaque. / Fujimoto, C.; Maeda, H.; Kokeguchi, S.; Takashiba, S.; Nishimura, Fusanori; Arai, H.; Fukui, K.; Murayama, Y.

In: Journal of Periodontal Research, Vol. 38, No. 4, 01.01.2003, p. 440-445.

Research output: Contribution to journalArticle

Fujimoto, C. ; Maeda, H. ; Kokeguchi, S. ; Takashiba, S. ; Nishimura, Fusanori ; Arai, H. ; Fukui, K. ; Murayama, Y. / Application of denaturing gradient gel electrophoresis (DGGE) to the analysis of microbial communities of subgingival plaque. In: Journal of Periodontal Research. 2003 ; Vol. 38, No. 4. pp. 440-445.
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