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Sensitive prion detection by in vitro amplification of abnormal prion protein

Research Project

Project/Area Number 20390287
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Infectious disease medicine
Research InstitutionNagasaki University

Principal Investigator

ATARASHI Ryuichiro  Nagasaki University, 大学院・医歯薬学総合研究科, 助教 (90452846)

Project Period (FY) 2008 – 2010
Project Status Completed (Fiscal Year 2010)
Budget Amount *help
¥18,460,000 (Direct Cost: ¥14,200,000、Indirect Cost: ¥4,260,000)
Fiscal Year 2010: ¥3,120,000 (Direct Cost: ¥2,400,000、Indirect Cost: ¥720,000)
Fiscal Year 2009: ¥3,120,000 (Direct Cost: ¥2,400,000、Indirect Cost: ¥720,000)
Fiscal Year 2008: ¥12,220,000 (Direct Cost: ¥9,400,000、Indirect Cost: ¥2,820,000)
Keywords感染症診断学 / プリオン病 / プリオン / 異常型プリオンタンパク / 試験管内増幅法 / リコンビナントタンパク / 早期診断
Research Abstract

The development of in vitro amplification technology for abnormal forms of prion protein (PrPSc) has generated the possibility for developing a novel diagnostic test for prion diseases. However, ultrasensitive PrPSc detection in easily accessible specimens such as cerebrospinal fluid (CSF) has not yet been successful in human prion diseases. In this study, we developed a new PrPSc amplification assay, designated "real-time QUIC (RT-QUIC)", allows for the detection of a minute amount of PrPSc in diluted sporadic Creutzfeldt?Jakob disease (sCJD) brain homogenate. Moreover, we assessed the technique first in a series of Japanese subjects and then in a blind study of 30 cerebrospinal fluid specimens from Australia, which achieved greater than 80% sensitivity and 100% specificity. These findings indicate the promising enhanced diagnostic capacity of RT-QUIC in the antemortem evaluation of suspected CJD.

Report

(4 results)
  • 2010 Annual Research Report   Final Research Report ( PDF )
  • 2009 Annual Research Report
  • 2008 Annual Research Report
  • Research Products

    (25 results)

All 2011 2010 2009 2008 Other

All Journal Article (13 results) (of which Peer Reviewed: 13 results) Presentation (9 results) Book (1 results) Remarks (2 results)

  • [Journal Article] Ultrasensitive human prion detection in cerebrospinal fluid by real-time quaking-induced conversion.2011

    • Author(s)
      Atarashi R, Satoh K, Sano K
    • Journal Title

      Nature Medicine 17

      Pages: 175-178

    • NAID

      120006984771

    • Related Report
      2010 Final Research Report
    • Peer Reviewed
  • [Journal Article] Ultrasensitive human prion detection in cerebrospinal fluid by real-time quaking-induced conversion.2011

    • Author(s)
      Atarashi R, Satoh K, Sano, K, (以下14名省略)
    • Journal Title

      Nature Medicine

      Volume: 17 Pages: 175-178

    • NAID

      120006984771

    • Related Report
      2010 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Mammalian prions generated from bacterially expressed prion protein in the absence of any mammalian cofactors.2011

    • Author(s)
      Kim JI, Cali I, Surewicz K, Kong Q, Raymond GJ, Atarashi R, (以下5名省略)
    • Journal Title

      J Biol Chem

      Volume: 285(19) Pages: 14083-14087

    • Related Report
      2010 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Rapid end-point quantitation of prion seeding activity with sensitivity comparable to bioassays.2010

    • Author(s)
      Wiham JM, Orru CD, Bessen RA, Atarashi R
    • Journal Title

      PLoS Pathogens 6(12)

    • Related Report
      2010 Final Research Report
    • Peer Reviewed
  • [Journal Article] Mammalian prions generated from bacterially expressed prion protein in the absence of any mammalian cofactors.2010

    • Author(s)
      Kim JI, Cali I, Surewicz K, Kong Q, Raymond GJ, Atarashi R
    • Journal Title

      J Biol Chem 285(19)

      Pages: 14083-14087

    • Related Report
      2010 Final Research Report
    • Peer Reviewed
  • [Journal Article] Rapid end-point quantitation of prion seeding activity with sensitivity comparable to bioassays.2010

    • Author(s)
      Wilham JM, Orru CD, Bessen RA, Atarashi R, (以下6名省略)
    • Journal Title

      PLoS Pathogens

      Volume: 6(12)

    • Related Report
      2010 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Distinct structures of scrapie prion protein (PrPSc)-seeded versus spontaneous recombinant prion protein fibrils revealed by hydrogen/deuterium exchange.2009

    • Author(s)
      Smirnovas V, Kim JI, Lu X, Atarashi R, Caughey B, Surewicz WK
    • Journal Title

      J Biol Chem 284(36)

      Pages: 24233-24241

    • Related Report
      2010 Final Research Report
    • Peer Reviewed
  • [Journal Article] Hyperefficient PrP Sc amplification of mouse-adapted BSE and scrapie strain by protein misfolding cyclic amplification technique.2009

    • Author(s)
      Fujihara A, Atarashi R
    • Journal Title

      FEBS J 276(10)

      Pages: 2841-2848

    • NAID

      120006984475

    • Related Report
      2010 Final Research Report
    • Peer Reviewed
  • [Journal Article] Distinct structures of scrapie prion protein(PrPSc)-seeded versus spontaneous recombinant prion protein fibrils revealed by hydrogen/deuterium exchange.2009

    • Author(s)
      Smirnovas V, Kim JI, Lu X, Atarashi R, Caughey B, Surewicz WK.
    • Journal Title

      J Biol Chem. 284(36)

      Pages: 24233-41

    • Related Report
      2009 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Hyperefficient PrP Sc amplification of mouse-adapted BSE and scrapie strain by protein misfolding cyclic amplification technique.2009

    • Author(s)
      Fujihara A, Atarashi R, Fuse T, Ubagai K, Nakagaki T, Yamaguchi N, Ishibashi D, Katamine S, Nishida N.
    • Journal Title

      FEBS J. 276(10)

      Pages: 2841-8

    • NAID

      120006984475

    • Related Report
      2009 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Recent advances in cell-free PrPSc amplification technique.2009

    • Author(s)
      Atarashi R
    • Journal Title

      Protein&peptide letters 16(3)

      Pages: 256-259

    • NAID

      120006982654

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Simplified ultrasensitive prion detection by recombinant PrP conversion with shaking.2008

    • Author(s)
      Atarashi R
    • Journal Title

      Nature Methods 5(3)

      Pages: 211-212

    • Related Report
      2010 Final Research Report
    • Peer Reviewed
  • [Journal Article] Simplified ultrasensitive prion detection by recombinant PrP conversion with shaking.2008

    • Author(s)
      Atarashi R, Wilham JM, Christensen L, Hughson AG, Moore RA, Johnson LM, Onwubiko HA, Priola SA, Caughey B.
    • Journal Title

      Nature methods 5(3)

      Pages: 211-212

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Presentation] シンポジウムI「プリオン病の疫学から治療まで」2010

    • Author(s)
      新竜一郎
    • Organizer
      第15回日本神経感染症学会
    • Place of Presentation
      福島
    • Year and Date
      2010-10-08
    • Related Report
      2010 Annual Research Report 2010 Final Research Report
  • [Presentation] Ultrasensitive human prion dectection in cerebrospinal fluids using real-time quaking-induced conversion.2010

    • Author(s)
      新竜一郎
    • Organizer
      Prion2010
    • Place of Presentation
      オーストリア(ザルツブルグ)
    • Year and Date
      2010-09-09
    • Related Report
      2010 Final Research Report
  • [Presentation] Ultrasensitive human prion detection in cerebrospinal fluid by real-time quaking-induced conversion.2010

    • Author(s)
      新竜一郎
    • Organizer
      Prion 2010
    • Place of Presentation
      オーストリア(ザルツブルグ)
    • Year and Date
      2010-09-09
    • Related Report
      2010 Annual Research Report
  • [Presentation] Ultrasensitive human prion dectection in cerebrospinal fluids using real-time quaking-induced conversion.2010

    • Author(s)
      新竜一郎
    • Organizer
      Asia-Oceania Symposium on Prion diseases 2010
    • Place of Presentation
      札幌
    • Year and Date
      2010-07-25
    • Related Report
      2010 Final Research Report
  • [Presentation] Ultrasensitive human prion detection in cerebrospinal fluid by real-time quaking-induced conversion.2010

    • Author(s)
      新竜一郎
    • Organizer
      Asia-Oceania Symposium on Prion diseases 2010
    • Place of Presentation
      札幌
    • Year and Date
      2010-07-05
    • Related Report
      2010 Annual Research Report
  • [Presentation] 正常型から異常型へのプリオンタンパク構造変換プロセスの解析2009

    • Author(s)
      新竜一郎
    • Organizer
      第82回日本生化学会
    • Place of Presentation
      神戸
    • Year and Date
      2009-10-22
    • Related Report
      2010 Final Research Report 2009 Annual Research Report
  • [Presentation] Real-time QUIC法によるクロイツフェルトヤコブ病患者由来髄液中のPrp^<Sc>の検出2009

    • Author(s)
      新竜一郎
    • Organizer
      2009年プリオン研究会
    • Place of Presentation
      宮城県刈田郡(ラフオーレ蔵王)
    • Year and Date
      2009-08-30
    • Related Report
      2010 Final Research Report
  • [Presentation] Real-time QUIC(QUaking-Induced Conversion)によるクロイツフェルトヤコブ病患者由来髄液中のPrP^<Sc>の検出2009

    • Author(s)
      新竜一郎
    • Organizer
      2009年プリオン研究会
    • Place of Presentation
      宮城県刈田郡
    • Year and Date
      2009-08-30
    • Related Report
      2009 Annual Research Report
  • [Presentation] PMCA法によるマウスプリオン株の高効率の増幅2008

    • Author(s)
      新 竜一郎
    • Organizer
      第56回日本ウイルス学会学術集会
    • Place of Presentation
      岡山コンベンションセンター
    • Year and Date
      2008-10-28
    • Related Report
      2008 Annual Research Report
  • [Book] プリオン病の髄液診断の可能性Annual Review神経2011

    • Author(s)
      新竜一郎、佐藤克也
    • Related Report
      2010 Final Research Report
  • [Remarks] ホームページ等

    • URL

      http://www.med.nagasaki-u.ac.jp/mmi/cmb/index.html

    • Related Report
      2010 Final Research Report
  • [Remarks]

    • URL

      http://www.mdp.nagasaki-u.ac.jp/ddid/staff/index.html

    • Related Report
      2008 Annual Research Report

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Published: 2008-04-01   Modified: 2016-04-21  

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