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Development of an ultrasensitive and specific dengue fever diagnostic system

Research Project

Project/Area Number 20H04556
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Review Section Basic Section 90130:Medical systems-related
Research InstitutionWaseda University

Principal Investigator

Ito Etsuro  早稲田大学, 教育・総合科学学術院, 教授 (80203131)

Co-Investigator(Kenkyū-buntansha) 戸谷 勇輝  早稲田大学, 教育・総合科学学術院, 助手 (90843897)
Project Period (FY) 2020-04-01 – 2023-03-31
Project Status Completed (Fiscal Year 2022)
Budget Amount *help
¥17,810,000 (Direct Cost: ¥13,700,000、Indirect Cost: ¥4,110,000)
Fiscal Year 2022: ¥5,590,000 (Direct Cost: ¥4,300,000、Indirect Cost: ¥1,290,000)
Fiscal Year 2021: ¥5,590,000 (Direct Cost: ¥4,300,000、Indirect Cost: ¥1,290,000)
Fiscal Year 2020: ¥6,630,000 (Direct Cost: ¥5,100,000、Indirect Cost: ¥1,530,000)
Keywordsタンパク質測定 / 感染症 / デング熱 / デング熱ウイルス / 早期診断 / 超高感度測定 / デングウイルス / 極微量タンパク質測定法
Outline of Research at the Start

我が国でも今後増加すると思われる熱帯性感染症などの早期診断を確実に行うためには、検体から極微量の病原体成分を短時間で特異的に検出する超高感度検出法を開発する必要がある。これまでは病原体の核酸増幅法が最高感度として推奨されてきたが、今回の目的にはさらなる高感度化が必要となる。我々は、サンドイッチELISA法とチオNADサイクリング法とを組み合わせた「極微量タンパク質測定法」の原理を発明した。本研究ではこの原理に基づいた測定法を最適化することで、タンパク質を超高感度で測定する世界初のシステムの構築を目指す。その適用例としてデング熱(デングウイルス)の早期診断システムの構築を試みる。

Outline of Final Research Achievements

To achieve early diagnosis of tropical infectious diseases, which are expected to increase in Japan in the future, it is necessary to develop an ultrasensitive detection method to specifically detect trace amounts of pathogen components from specimens in a short time. We have invented the principle of "ultra trace protein assay" combining the sandwich ELISA and thio-NAD cycling methods. By optimizing the assay based on this principle, we aimed to construct the system to measure proteins with ultrasensitivity. As an example of its application, we attempted to construct an early diagnosis system for dengue fever (dengue virus). The dengue virus NS1 protein was used as a model protein, and patient samples were transferred from Kaohsiung Medical University in Taiwan as a joint research project.

Academic Significance and Societal Importance of the Research Achievements

本研究によって超高感度タンパク質測定法が確立できたので、血中のタンパク質のみならず、さらに格段に低い濃度で存在すると考えられる尿中または唾液中のタンパク質の測定が可能となるはずである。これによって非侵襲的診断が可能となり、特に採血が困難な新生児や老齢の患者にとっては極めて安心・安全な診断法となる。この社会的意義は大きいと考える。なお本方法は、抗体を交換するだけでいかなる感染症にも応用でき、極めて高い汎用性がある点に、学術的意義がある。

Report

(4 results)
  • 2022 Annual Research Report   Final Research Report ( PDF )
  • 2021 Annual Research Report
  • 2020 Annual Research Report
  • Research Products

    (11 results)

All 2022 2021 Other

All Int'l Joint Research (1 results) Journal Article (10 results) (of which Int'l Joint Research: 4 results,  Peer Reviewed: 10 results,  Open Access: 10 results)

  • [Int'l Joint Research] 台湾・高雄医学大学(その他の国・地域)

    • Related Report
      2020 Annual Research Report
  • [Journal Article] Ultrasensitive ELISA detection of proteins in separated lumen and membrane fractions of cancer cell exosomes2022

    • Author(s)
      K.Iha, N.Tsurusawa, H.-Y.Tsai, M.-W.Lin, H.Sonoda, S.Watabe, T.Yoshimura, E.Ito
    • Journal Title

      Anal. Biochem.

      Volume: 654 Pages: 114831-114831

    • DOI

      10.1016/j.ab.2022.114831

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Ultrasensitive detection of GRP78 in exosomes and observation of migration and proliferation of cancer cells by application of GRP78-containing exosomes2022

    • Author(s)
      N.Tsurusawa, K.Iha, A.Sato, H.-Y.Tsai, H.Sonoda, S.Watabe, T.Yoshimura, D.-C.Wu, M.-W.Lin, E.Ito
    • Journal Title

      Cancers

      Volume: 14 Issue: 16 Pages: 3887-3887

    • DOI

      10.3390/cancers14163887

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Gastric cancer cell-derived exosomal GRP78 enhances angiogenesis upon stimulation of vascular endothelial cells2022

    • Author(s)
      K.Iha, A.Sato, H.-Y.Tsai, H.Sonoda, S.Watabe, T.Yoshimura, M.-W.Lin, E.Ito
    • Journal Title

      Curr. Issues Mol. Biol.

      Volume: 44 Issue: 12 Pages: 6145-6157

    • DOI

      10.3390/cimb44120419

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Non-amplification nucleic acid detection with thio-NAD cycling2022

    • Author(s)
      S.Yamura, N.Kawada, S.Yamakado, Y.Kyosei, S.Watabe, T.Yoshimura, Y.Murase, S.Mitarai, E.Ito
    • Journal Title

      J. Microbiol. Meth.

      Volume: 204 Pages: 106647-106647

    • DOI

      10.1016/j.mimet.2022.106647

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Zeptomole Detection of an Enzyme by a Simple Colorimetric Method2021

    • Author(s)
      Iha K, Kyosei Y, Namba M, Makioka D, Yamura S, Watabe S, Yoshimura T, Ito E.
    • Journal Title

      Analytical Sciences

      Volume: 37 Issue: 10 Pages: 1469-1472

    • DOI

      10.2116/analsci.21N009

    • NAID

      130008102515

    • ISSN
      0910-6340, 1348-2246
    • Year and Date
      2021-10-10
    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Improved Detection Sensitivity of an Antigen Test for SARS-CoV-2 Nucleocapsid Proteins with Thio-NAD Cycling2021

    • Author(s)
      Kyosei Y, Namba M, Yamura S, Watabe S, Yoshimura T, Sasaki T, Shioda T, Ito E.
    • Journal Title

      Biological and Pharmaceutical Bulletin

      Volume: 44 Issue: 9 Pages: 1332-1336

    • DOI

      10.1248/bpb.b21-00387

    • NAID

      130008082348

    • ISSN
      0918-6158, 1347-5215
    • Year and Date
      2021-09-01
    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Ultrasensitive Detection of SARS-CoV-2 Spike Proteins Using the Thio-NAD Cycling Reaction: A Preliminary Study before Clinical Trials2021

    • Author(s)
      Kyosei Y, Namba M, Makioka D, Kokubun A, Watabe S, Yoshimura T, Sasaki T, Shioda T, Ito E.
    • Journal Title

      Microorganisms

      Volume: 9 Issue: 11 Pages: 2214-2214

    • DOI

      10.3390/microorganisms9112214

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Modified ELISA for Ultrasensitive Diagnosis2021

    • Author(s)
      Tsurusawa N, Chang J, Namba M, Makioka D, Yamura S, Iha K, Kyosei Y, Watabe S, Yoshimura T, Ito E.
    • Journal Title

      J Clin Med.

      Volume: 10 Issue: 21 Pages: 5197-5197

    • DOI

      10.3390/jcm10215197

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Antigen tests for COVID-192021

    • Author(s)
      Kyosei Y, Yamura S, Namba M, Yoshimura T, Watabe S, Ito E.
    • Journal Title

      Biophysics and Physicobiology

      Volume: 18 Issue: 0 Pages: 28-39

    • DOI

      10.2142/biophysico.bppb-v18.004

    • NAID

      130008000162

    • ISSN
      2189-4779
    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Early diagnosis with ultrasensitive ELISA2021

    • Author(s)
      Ito E, Iha K, Yoshimura T, Nakaishi K, Watabe S.
    • Journal Title

      Adv Clin Chem.

      Volume: 101 Pages: 121-133

    • DOI

      10.1016/bs.acc.2020.06.002

    • ISBN
      9780128244159
    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access

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Published: 2020-04-28   Modified: 2024-01-30  

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