2002 Fiscal Year Final Research Report Summary
Structural analysis of oral microbiota by the molecular biology-technique, and its systematization
| Project/Area Number |
13672202
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Periodontal dentistry
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| Research Institution | RIKEN |
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Principal Investigator |
SAKAMOTO Mitsuo Molecular Microbial Functions Division , Contract Researcher, 微生物機能解析室, 協力研究員 (50321766)
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| Co-Investigator(Kenkyū-buntansha) |
UMEDA Makoto Department of Hard Tissue engineering, Graduate School,Tokyo Medical and Dental University , Assistant, 大学院・医歯学総合研究科, 助手 (90193937)
BENNO Yoshimi Molecular Microbial Functions Division , Head of Molecular Microbial Functions Division, 微生物機能解析室, 室長(副主任研究員待遇) (40087599)
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| Project Period (FY) |
2001 – 2002
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| Keywords | Periodontal disease / Oral bacteria / 16S rRNA gene / T-RFLP / Quantitative PCR |
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| Research Abstract |
Terminal restriction fragment length polymorphism (T-RFLP) analysis was applied to characterize oral bacterial flora in saliva from 18 healthy subjects and 18 patients with periodontitis. The 16S rRNA genes (rDNAs) of oral bacteria and spirochetes in saliva were amplified by PCR with a 6-FAM-labeled universal forward primer (27F) and a universal reverse primer (1492R) or the Spirochaeta-selective reverse primer. The 16S rDNAs were digested with restriction enzymes with 4-bp recognition sites (Hhal or MspI) and analyzed by using an automated DNA sequencer. T-RFLP patterns were numerically analyzed using a computer program. In the analysis of oral bacterial community, patterns derived from periodontally healthy subjects and patients with periodontitis were grouped into different clusters, though with some uncertainty. Samples from patients with periodontitis tended to cluster into their respective types (aggressive and chronic periodontitis), although this was not very clear. Analysis of spirochetal community using T-RFLP showed that the patterns derived from patients with periodontitis were grouped more as compared with the analysis of oral bacterial community. These results suggest that samples from patients with periodontitis contain an unexpected diversity. T-RFLP patterns of 16S rDNAs from saliva samples of two periodontally healthy subjects over a 5-week period showed host-specific relatively stable oral bacterial flora. Our study indicates that T-RFLP analysis is useful for the assessment of diversity of oral bacterial flora and rapid comparison of the community structure between subjects with and without periodontitis.
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Research Products
(2 results)
