Document Detail

Application of denaturing gradient gel electrophoresis (DGGE) to the analysis of microbial communities of subgingival plaque.
MedLine Citation:
PMID:  12828664     Owner:  NLM     Status:  MEDLINE    
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.
C Fujimoto; H Maeda; S Kokeguchi; S Takashiba; F Nishimura; H Arai; K Fukui; Y Murayama
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of periodontal research     Volume:  38     ISSN:  0022-3484     ISO Abbreviation:  J. Periodont. Res.     Publication Date:  2003 Aug 
Date Detail:
Created Date:  2003-06-27     Completed Date:  2003-10-02     Revised Date:  2008-11-21    
Medline Journal Info:
Nlm Unique ID:  0055107     Medline TA:  J Periodontal Res     Country:  Denmark    
Other Details:
Languages:  eng     Pagination:  440-5     Citation Subset:  D; IM    
Department of Patho-Physiology, Division of Periodontal Science, Okayama University Graduate School of Medicine and Dentistry, Okayama, Japan.
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MeSH Terms
5' Flanking Region / genetics
Actinobacillus actinomycetemcomitans / classification
Aggressive Periodontitis / microbiology
Bacteria / classification*,  genetics
DNA, Bacterial / genetics
Dental Plaque / microbiology*
Electrophoresis, Polyacrylamide Gel
GC Rich Sequence / genetics
Genome, Bacterial
Middle Aged
Neisseria / classification
Periodontal Pocket / microbiology
Polymerase Chain Reaction
Porphyromonas gingivalis / classification
Prevotella intermedia / classification
RNA, Ribosomal, 16S / genetics
Sequence Analysis, DNA
Streptococcus / classification
Reg. No./Substance:
0/DNA, Bacterial; 0/RNA, Ribosomal, 16S

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