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Development of sensitive diagnostic methods for transmissible spongiform encephalopathies.

Research Project

Project/Area Number 07556068
Research Category

Grant-in-Aid for Scientific Research (A)

Allocation TypeSingle-year Grants
Section展開研究
Research Field Applied veterinary science
Research InstitutionObihiro University of Agricutture and Veterinary Medicine

Principal Investigator

SHINAGAWA Morikazu  Obihiro Univ., Vet.Med., Prof., 畜産学部, 教授 (00001537)

Co-Investigator(Kenkyū-buntansha) KUBO Masanori  N.I.A.H.Lab.Pathol. Diag. Head Res., 家畜衛生試験場・病理診断室, 室長
YAMADA Haruo  Obihiro Univ., Vet.Med., Prof., 畜産学部, 教授 (20003113)
HIRUOKA Hidefumi  Obihiro Univ., Vet.Med., Ass.Prof., 畜産学部, 助教授 (60238665)
中川 迪夫  帯広畜産大学, 畜産学部, 教授 (60237237)
堀内 基広  帯広畜産大学, 原虫病分子免疫研究センター, 助教授 (30219216)
Project Period (FY) 1995 – 1997
Project Status Completed (Fiscal Year 1997)
Budget Amount *help
¥11,800,000 (Direct Cost: ¥11,800,000)
Fiscal Year 1997: ¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1996: ¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 1995: ¥6,300,000 (Direct Cost: ¥6,300,000)
KeywordsTransmissible spongiform encephalopathies / Preclinical diagnosis / ELISA / ELISA法 / 早期診断 / ウエスタンブロット
Research Abstract

In order to increase the sensitivity for detection of prion protein, the method to prepare samples from tissues for Western blotting was reformed. In mouse scrapie model, prion protein could be detected in the samples prepared from the spleen 1 week after intraperitoneal infection by the new method. It is to be desirable that more convenient method than Western blotting to examine many samples for the presence of prion protein. We developed an ELISA whose sensitivity was about two times higher than that of Western blotting using mouse model. We first confirmed that this ELISA method could be used for sheep scrapie samples, then applied the ELISA method for near 300 sheep samples obtained from slaughter house in Hokkaido.
We are developing the method for preparing samples to detect prion protein in contaminated collagen. We found the ways to reduce viscosity of collagen solution by enzymes and to concentrate prion from large volume of the enzyme treated collagen solution.
We also examined polymorphisms of sheep prion gene in relation to sheep with scrapie, expression of prion protein in various sheep organs, and polymorphisms of sheep apolipoprotein E gene as a risk factor of scrapie.

Report

(4 results)
  • 1997 Annual Research Report   Final Research Report Summary
  • 1996 Annual Research Report
  • 1995 Annual Research Report
  • Research Products

    (15 results)

All Other

All Publications (15 results)

  • [Publications] Ikeda, T.et.al.: "Amino acid polymorphisms of PrP with reference to onset of scrapie in sSuffolk and Corriedale sheep in Japan." J.Gen.Virol.76. 2577-2581 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Horiuchi, M.et.al.: "A cellular form of prion protein (PrPC) exists in many non-neuronal tissues of sheep." J.Gen.Virol.76. 2583-2587 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Grathwohl, K-U.et.al.: "Improvement of PrPSc-detection in mouse spleen early at the preclinical stage of scrapie with collagenase-completed---" Arch.Virol.141. 1863-1874 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Grathwohl, K-U.et.al.: "Sensitive enzyme-linked immunosorbent assay fordetection of PrPSc in crude tissue extracts from scrapie-affected--" J.Virol.Methods. 64. 205-216 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Komatsu, Y.et.al.: "Chracterization of the sheep apolipoprotein E(Apo E)gene and allelic variations of the Apo E gene in scrapie-----" Gene. (in press). (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Ikeda, T., Horiuchi, M., Ishiguro, N., Muramatsu, Y., Kai-Uwe, G.D., and Shinagawa, M.: "Amino acid polymorphisms of PrP with reference to onset of scrapie in Suffolk and Corriedale sheep in Japan." J.Gen. Virol.76. 2577-2581 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Horiuchi, M., Yamazaki, N., Ikeda, T., Ishiguro, N., and Shinagawa, M.: "A cellular form of prion protein (PrPC) exists in many non-neuronal tissues of sheep." J.Gen. Virol.76. 2583-2587 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Grathwohl, K-U.D.Horiuchi M., Ishiguro N.and Shinagawa, M.: "Improvement of PrPSc-detection in mouse spleen early at the preclinical stage of scrapie with collagenase-completed tiusse homogenization and Sarkosyl-NaCl extraction of PrPSc." Arch. Virol.141. 1863-1874 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Grathwohl K-U.D., Horiuchi, M., Ishiguro N.and Shinagawa, M.: "Sensitive enzyme-linked immunosorbent assay for detection of PrPSc in crude tissue extracts from scrapie-affected mice." J.Virol. Methods. 64. 205-216 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Komatsu, Y.Horiuchi, M., Ishiguro, N., Matsui, T.and Shinagawa, M.: "Characterization of the sheep apolipoprotein E (Apo E) gene and allelic variations of the ApoE vgene in scrapie Suffolk sheep." Gene.(in press). (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Grathwohl,K-U.et.al.: "Sensitive enzyme-Iinked immunosorbent assy for detection of PrPSc in crude tissue extracts from scrapie affected mice." J.Virol.Methods. 64. 205-216 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Komatsu,Y.et.al.: "Characterization of the sheep apolipoprotein E (Apo E) gene and allelic variation of the Apo E gene in scrapie----" Gene. in press.

    • Related Report
      1997 Annual Research Report
  • [Publications] K.W.D.Grathwol et al.: "Improvement of PrPSc-detection in mouse spleen early at the preclinica stage of scrapie with collagenase-completed tissuehomogenization and Sarkosyl-NaCl extraction of PrPSc" Arch.Virol.141. 1863-1874 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] S.Inoue et al.: "Characterization of the bovine prion protein gene : the expression requires interaction between the promter and intron." J.Vet.Med.Sci.59(in press). (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] K.-W.D.Grathwol et al.: "Sensitive enzyme-linked immunosor bent assay for detection of PrPSc in crude tissue extracts from scrapie-affected mice." J.Virol.Methods. (in press). (1997)

    • Related Report
      1996 Annual Research Report

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Published: 1995-04-01   Modified: 2016-04-21  

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