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Clinical application of serotyping of clinical isolates of Legionella pneumophila using MALDI-TOF MS

Research Project

Project/Area Number 19K07947
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Review Section Basic Section 52010:General internal medicine-related
Research InstitutionAzabu University

Principal Investigator

Sogawa Kazuyuki  麻布大学, 生命・環境科学部, 准教授 (50436440)

Co-Investigator(Kenkyū-buntansha) 古畑 勝則  麻布大学, 生命・環境科学部, 教授 (00308305)
谷口 俊文  千葉大学, 医学部附属病院, 講師 (20724826)
石毛 崇之  千葉大学, 医学部附属病院, 臨床検査技師 (30757315)
小寺 義男  北里大学, 理学部, 教授 (60265733)
Project Period (FY) 2019-04-01 – 2022-03-31
Project Status Completed (Fiscal Year 2021)
Budget Amount *help
¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2021: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2020: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2019: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Keywordsレジオネラ属菌 / 質量分析計 / 細菌同定 / 血清型別 / MALDI-TOF MS / 病原微生物
Outline of Research at the Start

レジオネラ肺炎による死亡数は依然として高く、呼吸不全を呈する市中肺炎の原因菌に限ると肺炎球菌(18.2%)に次いでレジオネラ菌(14.4%)が多い。レジオネラ肺炎の診断には尿中抗原検査が有用であるが、L. pneumophilia血清1型以外の感度は著しく低い。
本研究は質量分析計(MALDI-TOF MS)で同定率の低いレジオネラ属菌(L. pneumophilia, L. micdadei, L. bozemanii, L. dumoffii)に注目して、同定率の向上及びL. pneumophiliaの血清型別法を構築し、臨床応用を目指す。

Outline of Final Research Achievements

Legionella pneumophila (L. pneumophila) is responsible for most Legionnaire’s disease cases diagnosed worldwide. MALDI-TOF MS can be used to identify the L. pneumophila serogroup. In this study, we compared three sample preparation methods that are compatible with MALDI-TOF MS: DCTM, OTEM, and ITEM. The aim was to improve the low identification rates for L. pneumophila, and establish and validate a simple, rapid and robust MALDI-TOF MS-based method for routine use in microbiological laboratories for assignment of L. pneumophila isolates to serogroups and identification of reliable peak biomarkers. Using ITEM, 100.0% of hot spring water samples and clinical isolates were correctly identified at the species level. Augmented reference spectra correctly identified all 29 strains at the species level and 29 isolates at the serogroup level, displaying sensitivity, specificity and accuracy of 100.0% for serogroup assignment.

Academic Significance and Societal Importance of the Research Achievements

レジオネラ肺炎による死亡数は依然として高く、呼吸不全を呈する市中肺炎の原因菌に限ると肺炎球菌(18.2%)に次いでレジオネラ菌(14.4%)が多い。レジオネラ肺炎の診断には尿中抗原検査が有用であるが、L. pneumophilia血清1型以外の感度は著しく低い。質量分析計による細菌同定はサンプル調整が容易で、測定操作も簡便であり、一菌種約5分で同定結果が得られる。また、レジオネラ属菌の同定率の向上、同定及びL. pneumophiliaの血清型別に要する時間の短縮、ランニングコストが約1/10に減り、社会的意義は非常に高い。

Report

(4 results)
  • 2021 Annual Research Report   Final Research Report ( PDF )
  • 2020 Research-status Report
  • 2019 Research-status Report
  • Research Products

    (10 results)

All 2021 2020 2019

All Journal Article (7 results) (of which Peer Reviewed: 7 results,  Open Access: 5 results) Presentation (3 results) (of which Invited: 2 results)

  • [Journal Article] Examination of conditions for regular internal quality control in identification of microorganisms using MALDI-TOF MS2021

    • Author(s)
      SOGAWA KAZUYUKI、KIYOSUKE MAKIKO、HATTORI KANAKO、MURATA SYOTA、WATARI TOMOHISA、KUSABA KOJI、SAEKI YUJI、HORIUCHI KAZUKI、YASUDA KAZUNARI、UECHI KOHEI、OKUMURA HAJIME、SEKIGUCHI SACHIE、FUJINAGA AZUMI、MURAKAMI MASAMI、NAKAYAMA TOMOHIRO
    • Journal Title

      Biocontrol Science

      Volume: 26 Issue: 2 Pages: 99-104

    • DOI

      10.4265/bio.26.99

    • NAID

      130008048753

    • ISSN
      1342-4815, 1884-0205
    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Usefulness of the MALDI-TOF MS technology with membrane filter protocol for the rapid identification of microorganisms in perioperative drainage fluids of hepatobiliary pancreatic surgery2021

    • Author(s)
      Sogawa Kazuyuki、Takano Shigetsugu、Ishige Takayuki、Yoshitomi Hideyuki、Kagawa Shingo、Furukawa Katsunori、Takayashiki Tsukasa、Kuboki Satoshi、Nomura Fumio、Ohtsuka Masayuki
    • Journal Title

      PLOS ONE

      Volume: 16 Issue: 2 Pages: e0246002-e0246002

    • DOI

      10.1371/journal.pone.0246002

    • Related Report
      2021 Annual Research Report 2020 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Fucosylated C4b-binding protein α C chain, a novel serum biomarker that predicts lymph node metastasis in pancreatic ductal adenocarcinoma2020

    • Author(s)
      Sogawa Kazuyuki、Yamanaka Sakino、Takano Shigetsugu、Sasaki Kosuke、Miyahara Yoji、Furukawa Katsunori、Takayashiki Tsukasa、Kuboki Satoshi、Takizawa Hirotaka、Nomura Fumio、Ohtsuka Masayuki
    • Journal Title

      Oncology Letters

      Volume: 21 Issue: 2 Pages: 127-135

    • DOI

      10.3892/ol.2020.12388

    • Related Report
      2020 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] 質量分析装置Matrix-assisted laser desorption/ionization time-of-flight mass spectrometryの細菌同定における内部精度管理実施要領の検討2020

    • Author(s)
      曽川 一幸, 清祐 麻紀子, 服部 佳奈子, 村田 正太, 渡 智久, 奥村 元, 関口 幸恵, 藤永 あずみ, 中山 智祥
    • Journal Title

      医療検査と自動化

      Volume: 45 Pages: 255-259

    • Related Report
      2020 Research-status Report
    • Peer Reviewed
  • [Journal Article] Use of the MALDI-TOF MS for identification of microorganisms causing felis urine samples2020

    • Author(s)
      Maeda Hiroto、Ishige Takayuki、Watanabe Toshifumi、Sogawa Kazuyuki
    • Journal Title

      Electrophoresis Letters

      Volume: 64 Issue: 1 Pages: 11-13

    • DOI

      10.2198/electroph.64.11

    • NAID

      130007871311

    • ISSN
      2189-2628, 2189-2636
    • Related Report
      2020 Research-status Report
    • Peer Reviewed
  • [Journal Article] Examination of the fecal microbiota in dairy cows infected with bovine leukemia virus2020

    • Author(s)
      Uchiyama Jumpei、Murakami Hironobu、Sato Reiichiro、Mizukami Keijiro、Suzuki Takehito、Shima Ayaka、Ishihara Genki、Sogawa Kazuyuki、Sakaguchi Masahiro
    • Journal Title

      Veterinary Microbiology

      Volume: 240 Pages: 108547-108547

    • DOI

      10.1016/j.vetmic.2019.108547

    • Related Report
      2019 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] A point mutation to the long terminal repeat of bovine leukemia virus related to viral productivity and transmissibility2019

    • Author(s)
      Murakami Hironobu、Todaka Haruna、Uchiyama Jumpei、Sato Reiichiro、Sogawa Kazuyuki、Sakaguchi Masahiro、Tsukamoto Kenji
    • Journal Title

      Virology

      Volume: 537 Pages: 45-52

    • DOI

      10.1016/j.virol.2019.08.015

    • Related Report
      2019 Research-status Report
    • Peer Reviewed / Open Access
  • [Presentation] 質量分析装置Matrix-assisted laser desorption/ionization time-of-flight mass spectrometryの細菌同定における内部精度管理の検討2020

    • Author(s)
      曽川一幸、清祐麻紀子、服部佳奈子、村田正太、渡 智久、奥村 元、関口幸恵、藤永あずみ、中山智祥
    • Organizer
      第71回日本電気泳動学会総会
    • Related Report
      2020 Research-status Report
    • Invited
  • [Presentation] ラットの Clostridium difficile 感染モデルにおける腸内細菌叢に関する検討2020

    • Author(s)
      矢野彩音、石毛崇之、伊藤智、安達明、塚本篤士、石﨑直人、古畑勝則、曽川一幸
    • Organizer
      第39回アルコール医学生物学研究会学術集会
    • Related Report
      2019 Research-status Report
  • [Presentation] ラットのアルコール性肝障害モデルにおける血中及び腸内細菌叢の変化2019

    • Author(s)
      曽川一幸
    • Organizer
      第54回アルコール・アディクション医学会学術総会
    • Related Report
      2019 Research-status Report
    • Invited

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Published: 2019-04-18   Modified: 2023-01-30  

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