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The establishment of diagnostic tool for causative microorganisms using laser capture microdissection

Research Project

Project/Area Number 17K08988
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Research Field Laboratory medicine
Research InstitutionUniversity of Miyazaki

Principal Investigator

Ichiro Takajo  宮崎大学, 医学部, 准教授 (20418841)

Co-Investigator(Kenkyū-buntansha) 岡山 昭彦  宮崎大学, 医学部, 教授 (70204047)
越本 知大  宮崎大学, フロンティア科学総合研究センター, 教授 (70295210)
Project Period (FY) 2017-04-01 – 2020-03-31
Project Status Completed (Fiscal Year 2019)
Budget Amount *help
¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
Fiscal Year 2019: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2018: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2017: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Keywords細菌検査 / レーザーマイクロダイセクション / Laser Microdissection / 細菌同定 / 16S rRNA / 感染症 / 臨床検査医学
Outline of Final Research Achievements

To detect causative microorganisms is important to select accuracy treatments in daily clinical practice of infectious diseases. Gram staining is a common technique to estimate causative microorganisms. However, sometimes, it is difficult to distinguish bacterial contamination of clinical samples in this method. In the present study, we tried to establish a novel method to detect "true" causative microorganisms using laser capture microdissection (LCM). Bacteria-phagocytized neutrophils were targeted and were collected by LCM. After nucleic acid extraction from these neutrophils, the bacteria-specific 16S ribosomal RNA was analyzed. In peritonitis model mice, a causative microorganism was detected by this method. Therefore, LCM method may be a useful tool to detect causative microorganism which was phagocytized by neutrophils. In future study, it is necessary to evaluate the sensitivity and specificity of this method in clinical samples.

Academic Significance and Societal Importance of the Research Achievements

喀痰や尿などの臨床検体における起炎菌の同定は,抗菌薬の選択を適切に行うために重要であるが,日常診療で取り扱う臨床検体はしばしば起炎菌以外の細菌に汚染されており,判断が難しい場合がある.好中球に貪食された細菌は起炎菌と考えられるため,本研究ではこの様な好中球をレーザーキャプチャーマイクロダイセクション(LCM)法で効率的に回収し,起炎菌を同定する方法の開発を進めた.グラム染色とLCM法を組み合わせ,細菌を貪食した好中球の回収と細菌特異的な16SリボゾームRNA遺伝子の同定が可能となった.腹膜炎モデル動物においても本方法により起炎菌の同定が可能であり,今後,臨床検体での応用が期待される.

Report

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

    (3 results)

All 2020 2018 2017

All Journal Article (1 results) (of which Peer Reviewed: 1 results,  Open Access: 1 results) Presentation (2 results)

  • [Journal Article] Detection of microbial genes in a single leukocyte by polymerase chain reaction following laser capture microdissection2018

    • Author(s)
      Yamada Akiteru、Umeki Kazumi、Saeki Yuji、Hashikura Yuuki、Nomura Hajime、Yamamoto Ikuo、Umekita Kunihiko、Takajo Ichiro、Koshimoto Chihiro、Okayama Akihiko
    • Journal Title

      Journal of Microbiological Methods

      Volume: 155 Pages: 42-48

    • DOI

      10.1016/j.mimet.2018.11.005

    • Related Report
      2018 Research-status Report
    • Peer Reviewed / Open Access
  • [Presentation] グラム染色標本からの新たな菌種同定法の開発.2020

    • Author(s)
      山田明輝
    • Organizer
      第31会日本臨床微生物学会総会・学術集会
    • Related Report
      2019 Annual Research Report
  • [Presentation] グラム染色とLaser Micro Dissection法を組み合わせた核酸診断法.2017

    • Author(s)
      山田明輝,山本成郎,梅北邦彦,高城一郎,岡山昭彦.
    • Organizer
      第91回日本感染症学会総会・学術講演会.
    • Related Report
      2017 Research-status Report

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

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