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DEVELOPMENT OF A DIAGNISTIC KIT FOR RAPID EVALUATION OF CARIES ACTIVITY

Research Project

Project/Area Number 04557093
Research Category

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

Allocation TypeSingle-year Grants
Research Field 小児・社会系歯学
Research InstitutionTHE UNIVERSITY OF TOKUSHIMA

Principal Investigator

OTA Fusao  TOKUSHIMA UNIVERSITY,SCHOOL OF MEDICINE,PROFESSOR, 医学部, 教授 (90035478)

Co-Investigator(Kenkyū-buntansha) HIROTA Katsuhiko  TOKUSHIMA UNIVERSITY,SCHOOL OF DENTISTRY,RESEARCH ASSOCIATE, 歯学部, 助手 (60199130)
ONO Tsuneko  TOKUSHIMA UNIVERSITY,SCHOOL OF DENTISTRY,LECTURER, 歯学部, 講師 (40035514)
YAMATO Masayuki  TOKUSHIMA UNIVERSITY,SCHOOL OF MEDICINE,RESEARCH ASSOCIATE, 医学部, 助手 (90210492)
MINATO Yoshihiro  TOKUSHIMA UNIVERSITY,HEALTH SERVICE CENTRE,PROFESSOR, 保健管理センター, 教授 (00035768)
NISHINO Mizuho  TOKUSHIMA UNIVERSITY,SCHOOL OF DENTISTRY,PROFESSOR, 歯学部, 教授 (90029976)
竹岡 あや  徳島大学, 医学部, 教務員 (00116831)
安富 豊  徳島大学, 歯学部, 助手 (50240891)
宮崎 寿次  長瀬産業(株), 研究開発センター, 主任研究員
福井 公明  徳島大学, 歯学部, 教授 (40035407)
Project Period (FY) 1992 – 1994
Project Status Completed (Fiscal Year 1994)
Budget Amount *help
¥4,600,000 (Direct Cost: ¥4,600,000)
Fiscal Year 1994: ¥300,000 (Direct Cost: ¥300,000)
Fiscal Year 1993: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1992: ¥2,800,000 (Direct Cost: ¥2,800,000)
KeywordsDiagnostic kit / Streptococcus.mutans / Enzyme immunoassay / Monoclonal antibody / membrane filtre-EIA / Caries risk / Mutans group of streptococci / う蝕 / プロトタイプ / EIA / DMF / 酵素標識抗体 / 迅速診断法 / 血清型 / ハイブリドーマ / 遺伝子型別
Research Abstract

It is been known for long time that caries is produced primarily by infection with Streptococcus mutans. Since the bacteria group is an endogenous in the oral cavity of humans, no effective strategies have been found. The bacterial group multiply to affect teeth primarily in children with high risks at 2-10ages of life. A rapid quantification of the bacterial group is essential for and a key towards predicting the development of dental decay. However, no such methods have been developed. Ota et al recently succeeded in preparing several monoclonal antibodies specific for the members of mutans group of streptococci. They have also developed an enzyme immunoassay using these monoclonal antibodies, which can quantify the number of Streptococcus mutans in the oral cavity in nearly 60 min after collection of test samples from patients.
This study is aimed at developing a diagnostic kit for rapid quantification of Streptococcus mutans in the human oral cavity. Attempts were also made to prepa … More re more number of monoclonal antibodies and their specification.
Six batches of monoclonal antibodies with different specificity were produced. All batches were tested for their specificity against 4 strains of S.cricetus, 3 strains of S.rattus, 15 strains of S.mutans 5 strains of S.sobrinus and 4 strains of S.downei. Among them monoclonal antibodies F-89 was specific for Streptococcus mutans. The monoclonal antibody was purified by use of affinity chromatography and labelled with horse radish peroxidase. A new identification system(membrane filtre-EIA)using the labelled mAb f-89 was found to detect Streptococcus mutans at a level of about 10^4 cells/mg of dental plaque or washings of patients teeth. A prototype of a diagnostic kit using this identification system has been produced and tested for efficacy against samples of dental plaque taken mainly from young girls.
Results have shown that positive reactions were in good agreement with caries history of the subjects examined. A commercial model from the prototype was also manufactured and awaits further clinical trials. Less

Report

(4 results)
  • 1994 Annual Research Report   Final Research Report Summary
  • 1993 Annual Research Report
  • 1992 Annual Research Report
  • Research Products

    (37 results)

All Other

All Publications (37 results)

  • [Publications] Ota, F.,他: "Immunochemical study of polysaccharide antigen in St. sobrinus and St. downei with a cross-reactive monoclonal antibody." FEMS Immunol. & Med. Microbiol.,. 8. 213-218 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Ono, T.,他: "Detection of Streptococcus mutans by PCR amplification of spaP gene." J. Med. Microbiol.,. 41. 231-235 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Yamato, M.,他: "Monoclonal antibodies to spirosin of Yersinia enterocolitica and analysis of the localization of spirosome by use of them." Microbiol. Immunol.,. 38. 177-182 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] 岡田 要、他: "Streptococcus mutansによる実験腎炎" 日本小児腎臓病学会雑誌. 7. 82-85 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Ota, F.,他: "Immunochemical analysis and localization of a cell wall antigen of Streptococcus rattus with a specific monoclonal antibody." Zbl. Bakt.,. (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] 大和 正幸、他: "分子遺伝学的手法を利用したS.mutansの同定" 日本細菌学雑誌. 50(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] 高木 篤編、斉藤 肇、他著: "エッセンシャル微生物学" 医歯薬出版株式会社 (印刷中), 740 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] 大森 俊彦、太田 房雄、秋本 茂 著: "微生物学" 朝倉書店, 315 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Batoni, G.Ota, F., Ghelardi, G.Sensesi, S.Barnini, S., Freer, G., Hirota, K., Gabriele, M.Marcucci, M.and Campa, M.: "Epidemiological survey of Streptococcus mutans in a group of adult patients living in Pisa(Italy)" Eur.J.Epidemiol.238-242 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Yasutomi, Y., Matsuda, Y.Miyazaki, T,Nakamura, T.Hirota, K., Fukui, K., Ota, F., and M.Nishino: "Development of a new caries activity test using enzyme immunoassay system with monoclonal antibody." Japan J.Pedia.Dent.30. 186-193 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Hirota, K., Ono, T., Fukui, K., Yamashita, K., Yamato, M., Ota, F., and Y.Minato.: "Immunological analysis of a cell surface antigen by monoclonal antibody f-77 in Streptococcus mutans and Streptococcus Pyogenes cells." Shikoku Acta Medica. 49. 8-15 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Hirota, K.Fernandez E., Ono., T., Miyazaki, T., Matsuda, Y.Nakamura, T.Yasutomi, Y., Nishino, M., Minato, Y.Ota, F.and K.Fukui.: "Simple and rapid identification of the mutans group of streptococci by enzyme immunoassay using monoclonal antibodies." Shikoku Acta Dent.5. 65-74 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Batoni, G., Marchetti, M., Ota, F., Ghelardi, E., Barnini, S., Inoue, H., Uchiyama, C., Hirota, K., Minato, Y., Guica, M.R., Gabriele, M., Campa, M.Senesi, S.: "First characterization in Italy of clinical isolates of mutans streptococci by using specific monoclonal antibodies." Eur.J.Epidemiol.9. 483-488 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Ota, F., Kato, H., Hirota, K., Inoue, H., Tomotake, H., Uchiyama, C., and Minato, Y.: "Immunochemical study of polysaccharide antigen in Streptococcus sobrinus and Streptococcus downei with a cross reactive monoclonal antibody." FEMS Immunol.Med.Microbiol.8. 213-218 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Fusao Ota, Hirohisa Kato, Katsuhiko Hirota, Susumu Imai, Hiroyuki Ishikawa, Masayuki Yamato and Yoshihiro Minato.: "Immunochemical analysis and localization of a cell wall antigen of Streptococcus rattus with a specific monoclonal antibody." Zbl.Bakt.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] Ota,F.,他: "Immunochemical study of polysaccharide antigen in St.sobrinus and St.downei with a cross-reactive monoclonal antibody." FEMS Immunol.& Med.Microbiol.,. 8. 213-218 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Ono,T.,他: "Detection of Streptococcus mutans by PCR amplification of spaP gene." J.Med.Microbiol.,. 41. 231-235 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Yamato,M.,他: "Monoclonal antibodies to spirosin of Yersinia enterocolitica and analysis of the localization of spiromome by use of them." Microbiol.Immunol.,. 38. 177-182 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] 岡田 要、他: "Streptococcus mutansによる実験腎炎" 日本小児腎臓病学会雑誌. 7. 82-85 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Ota,F.,他: "Immunochemical analysis and localization of a cell wall antigen of Streptococcus rattus with a specific monoclonal antibody." Zbl.Bakt.,. (印刷中). (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] 大和 正幸、他: "分子遺伝学的手法を利用したS.mutansの同定" 日本細菌学雑誌. 50(印刷中). (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] 高木 篤編、斉藤 肇、他著: "エッセンシャル微生物学" 医歯薬出版株式会社(印刷中), 740 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] 大森 俊彦、太田 房雄、秋本 茂、著: "微生物学" 朝倉書店, 315 (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] Ota,F.,他: "Immunochemical study of polysaccharide antigen in St.sobrinus and St.downei with a cross-reactive monoclonal antibody." FEMS Immunol.& Med.Microbiol.(印刷中). (1994)

    • Related Report
      1993 Annual Research Report
  • [Publications] Hirota,K.,他: "A cross-reactive polysaccharide antigen between Streptococcus mutans and Streptococcus pyogenes." Zbl.Bakt.Suppl.(印刷中). (1994)

    • Related Report
      1993 Annual Research Report
  • [Publications] Hirota,K.,他: "Streptococcus mutans has a polysaccharide antigenic determinant in common with Streptococcus porcinus." Zbl.Bakt.Suppl.(印刷中). (1994)

    • Related Report
      1993 Annual Research Report
  • [Publications] Marchetti,F.,他: "Incidence of mutans streptococci in insulin-dependent Diabetes mellitus patients." Zbl.Bakt.Suppl.(印刷中). (1994)

    • Related Report
      1993 Annual Research Report
  • [Publications] Batoni,G.,他: "First characterization in Italy of clinical isolates of mutans streptococci by using specific monoclonal antibodies." Eur.J.Epidemiol.(印刷中). (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] 弘田 克彦、他: "モノクロン抗体によるStreptococcus mutansおよびStreptococcus pyogenesの菌体表層抗原の免疫学的解析" 四国医学雑誌. 49. 8-15 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] 高木 篤編、斉藤 肇、他著: "エッセンシャル微生物学" 医歯薬出版株式会社 (印刷中), 740 (1994)

    • Related Report
      1993 Annual Research Report
  • [Publications] Batoni,G.,Ota,F.,: "Epidemiological survey of Streptococcus mutans in a group of adult patients living in Pisa(Italy)." Eur.J.Epidemiol.8. 238-242 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Ota,F.,Batoni,G.,: "Serological identification of mutans streptococci using MAbs in comparison with biochemical and DNA-DNA hybridisation techniques." Zbl.Bakt.(suppl.). 22. 84-87 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Hirota,K.,: "A comparison of various EIA plates for the analysis of polysaccharide antigens and serological identification of oral streptococci." Zbl.Bakt.(suppl.). 22. 116-118 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Nagamune,H.,Ota,F.,: "Monoclonal antibodies specific for a polysaccharide antigenic determinants of Streptococcus mutans(sorotypes c,e and f)." Zbl.Bakt.(suppl.). 22. 223-225 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] 安富 豊、松田 洋子,他: "単クロン抗体を用いた酵素抗体法の齲蝕活動試験への応用(第一報)" 小児歯科学雑誌. 30. 186-193 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] 弘田 克彦,他: "モノクロン抗体を使用した酵素免疫測定法によるミュータンスレンサ球菌の迅速簡易同定法" 四国歯学会雑誌. 5. 65-74 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] 高野 篤 編,斎藤 肇,他著: "エッセンシャル微生物学第III版" 医歯薬出版株式会社, 740 (1993)

    • Related Report
      1992 Annual Research Report

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Published: 1992-04-01   Modified: 2016-04-21  

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