| Project/Area Number |
13671913
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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 |
Morphological basic dentistry
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| Research Institution | Tokyo Dental College |
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Principal Investigator |
MIURA Tadashi Tokyo Dental College, Department of Dentistry, Assistant Professor, 歯学部, 助教授 (10266570)
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| Co-Investigator(Kenkyū-buntansha) |
YAMANAKA Ayumi Tokyo Dental College, Department of Dentistry, Assistant, 歯学部, 助手 (40231667)
ISHIHARA Kazuyuki Tokyo Dental College, Department of Dentistry, Assistant Professor, 歯学部, 助教授 (00212910)
KATO Tetsuo Tokyo Dental College, Department of Dentistry, Assistant Professor, 歯学部, 助教授 (00159253)
KIMIZUKA Ryuta Tokyo Dental College, Department of Dentistry, Assistant, 歯学部, 助手 (90287178)
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| Project Period (FY) |
2001 – 2003
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| Project Status |
Completed (Fiscal Year 2003)
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| Budget Amount *help |
¥2,500,000 (Direct Cost: ¥2,500,000)
Fiscal Year 2003: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 2002: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 2001: ¥900,000 (Direct Cost: ¥900,000)
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| Keywords | Campylobacter rectus / monoclonal antibody / Infection / Bacteroides forsythus / cloning / Periodontal bacteria / Periodontal lesion / detection / Bacteroides forsythus / C.rectus / monoclonal antibody / cloning / 嫌気性細菌 |
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| Research Abstract |
In the diagnosis of periodontal disease, microbial examinations can identify patients and periodontal sites that are undergoing active tissue destruction and can aid in understanding of fluctuations of periodontopathogens after treatment. Porphyromonas gingivalis, Actinobacillus actinornycetenicomitans, Fusobacterium nucleatum Bacteroides forsythus, Carnpylobacter rectus are known as major periodontal bacteria. Campylobacterrectushas often been detected in large numbers in deeper subgingival pockets and has been implicated in adult periodontitis and rapidly advancing periodontitis. It is gram-negative, microaerophilic bacterium, a few reports for clinical status and C.rectus were submitted. Therefore, the aim of this study is to clarify its pathogens.
1) We cultured successfully a great deal of Carnpylobacter rectus cells as a result of reforming a broth.
2) We prepared monoclonal antibodies and polyclonal ones. Campylobacter rectus ATCC 33238 whole cells were used as the immunogen.
3) We isolated three monoclonal antibodies specific to C.rectus. These antibodies recognized a peculiar 150 KDa protein of C.rectus, thought a S-layer. One of the antibodies did not cross-react to some clinical strains, showed that the S-layer has heterogeneity.
4) As a result that relationship between PD and the detection rate of C.rectus using dot-blot analysis was statistically significant. As periodontal pockets became deeper, the detection rate of C.rectus increased.
5) With one of the polyclonal antibodies, we made an attempt of cloning C.rectus genes from chromosomal DNA fragments treated by restriction enzyme. As a result, we got positive clones. We determined its DNA sequences of one of the clones. This gene had a specific sequence, and not homologous to known genes.
6) Furthermore in order to clarify C.rectus pathogenecities, we are planning to continue gene-cloning, and simultaneously we analyse Bacteroides forsythus cell surface antigens using several monoclonal antibodies.
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