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Development and clinical application of detection method of periodontopathic bacteria by using a non-radioactive DNA probe

Research Project

Project/Area Number 03557081
Research Category

Grant-in-Aid for Developmental Scientific Research (B)

Allocation TypeSingle-year Grants
Research Field Conservative dentistry
Research InstitutionTokyo Medical and Dental University

Principal Investigator

ISHIKAWA Isao  Tokyo Medical and Dental University Faculty of Dentistry Professor, 歯学部, 教授 (10014151)

Co-Investigator(Kenkyū-buntansha) UMEDA Makoto  Tokyo Medical and Dental University Faculty of Dentistry Research Assistant, 歯学部, 助手 (90193937)
安井 聡志  東京医科歯科大学, 歯科部, 助手 (40221642)
Project Period (FY) 1991 – 1993
Project Status Completed (Fiscal Year 1993)
Budget Amount *help
¥10,800,000 (Direct Cost: ¥10,800,000)
Fiscal Year 1993: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1992: ¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 1991: ¥6,000,000 (Direct Cost: ¥6,000,000)
Keywordsperiodontal diseases / periodontopathic bacteria / non-radioactive DNA probes / bacterial identification / 歯周病 / 組菌同定 / Porphyromonas gingivalis / 非放射性 / 血清中抗体価 / DNAプロ-ブ / 歯周病原細菌
Research Abstract

A dot blot DNA hybridization assay for a rapid identification of periodontopathic bacteria in subgingival plaque samples was developed by using bisulfite modified whole genomic DNA probe from which was immunoenzymatically visualized by anti SO_3^- antibody conjugated with alkalinephosphatase. Bisulfite modified DNA probe of P.gingivalis and P.intermedia showed high specificity and sensitivity to detect reference strains. The probe from P.gingivalis was applied on clinical subgingival plaque samples to evaluate the distribution of P.gingivalis in the dentition, and to compare the presence of P.gingivalis with clinical parameters. Fifteen adult periodontitis patients and 6 healthy volunteers were examined. Subgingival plaque samples were taken from 4 sites of all the remaining teeth. At the same time, probing depth and bleeding on probing (BOP) were also recorded. The detection percentage and amounts of P.gingivalis present were statistically compared with probing depth and BOP in the patient group. As a result, P.gingivalis was detected in all patients examined and 3 out of 6 healthy individuals. The detection average was 31% in patient group. When the probing depth was over 4 mm or BOP was positive, the detection percentage of P.gingivalis significantly increased. As more P.gingivalis was identified, the percentage of sites with deep probing depth or that were BOP positive increased significantly. However, P.gingivalis was also detected in clinically healthy sites, and P.gingivalis negative sites with deep probing depth or that were BOP positive were existed in the same patient. These results indicate that P.gingivalis play an important role, but is not the only microorganism responsible for adult periodontitis. Moreover, using RFLPs analysis we examined P.gingivalis strains including clinical isolates on their fimbrillin gene locus. The results indicate that genetic heterogeneity seems to be existing within the fimbrillin gene locus.

Report

(4 results)
  • 1993 Annual Research Report   Final Research Report Summary
  • 1992 Annual Research Report
  • 1991 Annual Research Report
  • Research Products

    (15 results)

All Other

All Publications (15 results)

  • [Publications] Yasui,S.et al: "Rapid identification of Porphyromonas gingivalis by bisulfite modified DNA probe method." Journal of Periodontal Research. 28. 98-101 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Zhang,Y.G.: "Heterogeniety of Porphyromonas gingivalis strains on fimbrillin gene locus by restriction fragment length polymorphism analysis." The Bulletin of Tokyo Medical and Dental University. 40. 113-123 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Kojima,T.et al: "Distribution of Porphyromonas gingivalis in adult periodontitis patients." Journal of Periodontology. 64. 1231-1237 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] 小島丈尚: "非放射性DNAプローブ法を用いた口腔内におけるPorphyromonas gingivalisの分布に関する研究" 口腔病学会雑誌. 61. 71-81 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Yasui, S., Kojima, T., Hata, S., Zhang, Y.G., Umeda, M.and Ishikawa, I.: "Rapid identification of Porphyromonas gingivalis by bisulfite modified DNA probe method." Journal of Periodontal Research. 28. 98-101 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Zhang, Y.G.: "Heterogeniety of Porphyromonas gingivalis strains on fimbrillin gene locus by restriction fragment length polymorphism analysis." The Bulletin of Tokyo Medical and Dental University. 40(3). 113-123 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Kojima, T., Yasui, S.and Ishikawa, I.: "Distribution of Porphyromonas gingivalis in adult periodontitis patients." Journal of Periodontology. 64. 1231-1237 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Kojima, T.: "A Study on distribution of Porphyromonas gingivalis in oral cavity by using a non-radioactive DNA probe method." The Journal of the Stomatological Society, Japan. 61. 71-81 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] 張 〓〓ら: "サザンブロット法を用いたPorphyromonas gingivalis菌フィンブリエ遺伝子の解析" 日本歯科保存学雑誌. 35. 81- (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] 小島 丈尚ら: "同一患者におけるPorphyromonas gingivalisの存在と血清中抗体価に関する研究" 日本歯科保存学雑誌. 35. 82- (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] S.YASUI et.al.: "Rapid identification of Porphyromonas gingivalis by bisulfite-modified DNA probe methocl." Journal of Perioclontal Research.

    • Related Report
      1992 Annual Research Report
  • [Publications] 小島 丈尚ら: "Porphyromonas gingivalisの分布と歯周疾患の臨床症状との関連性" 日本細菌学雑誌. 48. 269- (1993)

    • Related Report
      1992 Annual Research Report
  • [Publications] 石川 烈: "長寿科学総合研究平成三年度研究報告Vol.2ー高齢者の歯周疾患の病態解析及び診断法開発に関する研究ー" 長寿科学総合研究費中央事務局, 4 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Takehisa Kojima: "Identification of B.gingivalis with bisulfite modified DNA probe" Journal of Dental Research. 70. 505- (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] 石川 烈: "歯周疾患の免疫学的診断法及び非放射性DNA probeの確立による治療法の決定" 長寿科学総合研究. 2. 484-486 (1991)

    • Related Report
      1991 Annual Research Report

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Published: 1991-04-01   Modified: 2016-04-21  

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