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Establishment of highly sensitive ELISA to determine specific IgM antibody for diagnosis of acute cat scratch disease and elucidation of its pathology

Research Project

Project/Area Number 17K09013
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Research Field Laboratory medicine
Research InstitutionYamaguchi University

Principal Investigator

TSUNEOKA Hidehiro  山口大学, 大学院医学系研究科, 教授(特命) (40437629)

Co-Investigator(Kenkyū-buntansha) 大津山 賢一郎  山口大学, 大学院医学系研究科, 助教 (10432741)
徳田 信子  獨協医科大学, 医学部, 教授 (70227578)
Project Period (FY) 2017-04-01 – 2020-03-31
Project Status Completed (Fiscal Year 2019)
Budget Amount *help
¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000)
Fiscal Year 2019: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2018: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2017: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Keywords猫ひっかき病 / Bartonella henselae / ELISA-IgM / ELISA / ウエスタンブロット / IgM-WB / ウエスタンブロット法 / IgM-ELISA
Outline of Final Research Achievements

This study aimed to establish a highly sensitive IgM ELISA using recombinant protein antigens of Bartonella henselae for serodiagnosis of cat scratch disease (CSD). We used seven types of candidate genus as antigens found at 8-10kDa of the major protein in the B. henselae IgM antibody-positive patients and examined the antigenicity of these proteins by two methods: cell-free protein synthesis and protein synthesis introduced into Escherichia coli.
As a result, genus protein expression was confirmed in all the genera in both methods, while responsiveness to CSD patients’ sera was not confirmed and no antigenicity was clarified in the Western blot assay. Further investigation will be required to establish the optimal IgM ELISA for the detection of CSD.

Academic Significance and Societal Importance of the Research Achievements

猫ひっかき病(CSD)の病原菌Bartonella henselae は培養困難なため、血清学的診断法が有用であり高感度ELISAの開発が求められている。本研究は未だ存在しない“特異性の高い組換え蛋白質抗原を使用した高感度ELISA-IgM抗体価測定法の確立”を目指し、研究代表者らの一連の研究よりB. henselae菌体の8-10kDaの解析から研究を進めた。その結果、遺伝子解明とその組換え蛋白質生成合成はできたものの、それら蛋白質の抗原性は認められず、更なる検討が求められた。本ELISA確立時には一層のCSDの診断率向上と病態の解明が期待される。

Report

(4 results)
  • 2019 Annual Research Report   Final Research Report ( PDF )
  • 2018 Research-status Report
  • 2017 Research-status Report
  • Research Products

    (6 results)

All 2019 2018 2017

All Journal Article (4 results) (of which Int'l Joint Research: 2 results,  Peer Reviewed: 4 results,  Open Access: 3 results) Presentation (2 results)

  • [Journal Article] A CASE OF CAT SCRATCH DISEASE DIAGNOSED BY INDIRECT FLUORESCENT ANTIBODY ASSAY OF IGM SPECIFIC FOR A JAPANESE STRAIN OF Bartonella henselae2019

    • Author(s)
      Uchi Sho-Hei、Yanai Ryoji、Tsuneoka Hidehiro、Otsuyama Ken-ichiro、Sonoda Koh-Hei、Kimura Kazuhiro
    • Journal Title

      RETINAL Cases & Brief Reports

      Volume: Publish Ahead of Print Issue: 5 Pages: 1-1

    • DOI

      10.1097/icb.0000000000000854

    • Related Report
      2019 Annual Research Report 2018 Research-status Report
    • Peer Reviewed
  • [Journal Article] aCL/β2GPI and aPS/PT show synergic thrombogenic effects in suppressing anticoagulant activity of APC and stimulating tissue factor expression and TNF-α secretion by mononuclear cells2019

    • Author(s)
      Kaneshige Risa、Nojima Junzo、Motoki Yukari、Tsuneoka Hidehiro
    • Journal Title

      Thrombosis Research

      Volume: 181 Pages: 52-58

    • DOI

      10.1016/j.thromres.2019.07.006

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Bartonella henselae DNA in Seronegative Patients with Cat-Scratch Disease2018

    • Author(s)
      M.Yanagihara, H.Tsuneoka, A.Tanimoto, K.Otsuyama, J.Nishikawa, T.Matsui, J.Nojima, K.Ichihara
    • Journal Title

      Emerg Infect Dis

      Volume: 24 Issue: 5 Pages: 924-925

    • DOI

      10.3201/eid2405.152033

    • Related Report
      2018 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Utility of Bartonella henselae IgM Western Blot Bands for Serodiagnosis of Cat Scratch Disease2017

    • Author(s)
      Otsuyama Ken-ichiro、Tsuneoka Hidehiro、Yoshidomi Hiroka、Haraguchi Mio、Yanagihara Masashi、Tokuda Nobuko、Nojima Junzo、Ichihara Kiyoshi
    • Journal Title

      Journal of Clinical Microbiology

      Volume: 56 Issue: 1 Pages: 1-6

    • DOI

      10.1128/jcm.01322-17

    • Related Report
      2017 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Presentation] Bartonella henselae IgG-ELISAの培地別・精製法別抗原の比較検討2019

    • Author(s)
      大津山賢一郎・常岡英弘
    • Organizer
      第89回日本感染症学会西日本地方学術集会 浜松
    • Related Report
      2019 Annual Research Report
  • [Presentation] 猫ひっかき病血清学的診断におけるBartonella henselaeウエスタンブロット-IgM解析の有用性2018

    • Author(s)
      大津山賢一郎
    • Organizer
      第88回日本感染症学会西日本地方学術集会
    • Related Report
      2018 Research-status Report

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Published: 2017-04-28   Modified: 2021-02-19  

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