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Development of high-sensitive periodontitis diagnosis method using biotinylated polysaccharide antigens

Research Project

Project/Area Number 05454563
Research Category

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

Allocation TypeSingle-year Grants
Research Field 矯正・小児・社会系歯学
Research InstitutionKYUSHU UNIVERSITY

Principal Investigator

KOGA Toshihiko  Kyushu University, Faculty of Dentistry, Professor, 歯学部, 教授 (10037541)

Co-Investigator(Kenkyū-buntansha) YAMAGUCHI Noboru  Kyushu University, Faculty of Dentistry, Instructor, 歯学部, 助手 (00230368)
Project Period (FY) 1993 – 1994
Project Status Completed (Fiscal Year 1994)
Budget Amount *help
¥6,600,000 (Direct Cost: ¥6,600,000)
Fiscal Year 1994: ¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1993: ¥4,300,000 (Direct Cost: ¥4,300,000)
KeywordsPeriodontal disease / Polysaccharide antigen / Diagnosis method / Periodontopathic bacteria / Biotin / ELISA / 単球 / サイトカイン
Research Abstract

Periodontitis has been recently considered to be an infectious disease that is induced by several specific bacteria. In this study, we developed a rapid and specific method for measuring antibodies to periodontopathic bacteria. Serotype-specific polysaccharide antigens (SPAs) were extracted from whole cells of Actinobacillus actinomycetemcomitans by autoclaving and purified by chromatography. The purified SPAs were activated with cyanogen bromide and then treated with adipic acid dihydrazide. The preparations were coupled with biotin or bovine serum albumin (BSA) by the carbodiimide method. Enzyme-linked immunosorbent assay plates pretreated with avidin were allowed to react with biotinylated SPAs. Studies using rabbit anti-A.a.sera showed the assay utilizing avidin-biotin was very specific and sensitive. Antibodies to the antigens in 1 : 6,44 to 1 : 12,800 dilutions of human sera from subjects with periodontitis could be detected by using the assay. Subcutaneous immunization of rabbits with BSA-SPA induced high titers of serum anti-SPA antibodies, and these antibodies enhanced phagocytosis and killing of A.a.by human polymorphonuclear leukocytes. These findings suggest that proteinconjugated polysaccharide antigens may be useful for diagnosis of periodontal disease and elucidating an etiological role of the antigens in the induction of the infectious disease.

Report

(3 results)
  • 1994 Annual Research Report   Final Research Report Summary
  • 1993 Annual Research Report
  • Research Products

    (14 results)

All Other

All Publications (14 results)

  • [Publications] T.Nakano 他4名: "Monoclonal antibodies to the large chondroitin sulphate proteoglycan from bovine temporomandibular joint disc." Matrix. 13. 243-254 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] T.Koga 他6名: "The 64-kDa GroEL-like protein of Actinobacillus actinomycetemcomitans" Journal of Periodontal Research. 28. 47-477 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] T.Nishihara 他6名: "Membran-associated interleukin-1 promotes osteoclast-like cell formation in vivo" Bone Mineral. 25. 15-24 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Y.Nakano 他7名: "Molecular and immunological characterization of a 64-kDa protein of Actinobacillus actinomycetemcomitans" Oral Microbiology and Immunology. (印刷中). (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] M.Kawasaki 他5名: "An enzyme-linked immunosorbent assay for measuring antibodies to serotype-specific polysaccharide antigens of Actinobacillus actinomycetemcomitans" Journal of Microbiological Methods. (印刷中). (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] T.Nakano et al.: "Monoclonal antibodies to the large chondroitin sulphate proteoglycan from bovine temporomandibular joint disc." Matrix. 13. 243-254 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] T.Koga et al.: "The 64-kDa GroEL-like protein of Actinobacillus actinomycetemcomitans." J.Periodont.Res.28. 475-477 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] T.Nishihara et al.: "Membrane-associated interleukin-1 promotes osteoclast-like cell formation in vivo. Bone Mineral 25 : 15-24,1994." Bone Mineral. 25. 15-24 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Y.Nakano et al.: "Molecular and immunological characterization of a 64-kDa protein of Actinobacillus actinomycetemcomitans." Oral Microbiol.Immunol.(in press). (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] M.Kawasaki et al.: "An enzyme-linked immunosorbent assay for measuring antibodies to serotype-specific polysaccharide antigens of Actinobacillus actinomycetemcomitans." J.Microbiol.Methods. (in press). (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] T.Nishihara: "Membrane-associated interleukin-1 Promotes Osteoclast-like cell formation in vitro" Bone Mineral. 25. 15-24 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] M.Kawasaki: "An encyme-linked immunosorbent assay for measuring antibodies to actinomycetemcomitans" Journal of Microbiological Methods. (印刷中). (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] Y.Nakano: "Molecular and immunological characterization ob a 64-k Da Protein ob Actinobacillus actinomycetemcomitans" Oral Microbiology and Immunology. (印刷中). (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] Toshihiko Koga: "The 64-kDa GroEL-like protein of Actimobacillus actinomycetemcomitans." Journal of Periodontal Research. 28. 475-477 (1993)

    • Related Report
      1993 Annual Research Report

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Published: 1993-04-01   Modified: 2016-04-21  

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