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Comprehensive analysis of the microflora using a novel portable sequencer MinION

Research Project

Project/Area Number 17K10029
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Research Field Infectious disease medicine
Research InstitutionSaitama Medical University

Principal Investigator

Takuya Maeda  埼玉医科大学, 医学部, 教授 (20383763)

Co-Investigator(Kenkyū-buntansha) 各務 博  埼玉医科大学, 医学部, 教授 (30418686)
鈴木 穣  東京大学, 大学院新領域創成科学研究科, 教授 (40323646)
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: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2018: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2017: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Keywordsナノポアシーケンサー / MinION / 微生物叢 / 非結核性抗酸菌 / 核酸精製 / ストレプトコッカス / 下気道細菌叢 / ナノポア型シーケンサー / 多剤耐性結核 / 感染症 / 微生物
Outline of Final Research Achievements

In this study, we aimed to develop a technology for comprehensive and quantitative detection of various pathogens in the lung microenvironment. Particularly, we used nanopore sequencing technology, rather than the conventional culture-based method, using extracting and purifying all DNA from the collected clinical specimens. This method has the advantage that computer analysis of the obtained sequence reads allows simultaneous elucidation of different pathogens and their microbiota. In this study, we have characterized various pathogens present in the lung microenvironment, established a comprehensive protocol to extract and purify various pathogens derived from clinical specimens, and established a sequence analysis method using a nanopore sequencer, MinION.

Academic Significance and Societal Importance of the Research Achievements

本研究の成果は、国内で増加する非結核性抗酸菌症の診断と病態解明に活用できる検査法の確立にとどまらず、さまざまな感染症診断に展開できる新しい微生物同定技術へと発展できる可能性がある。この成果により、これまで診断が困難であった病原微生物の感染を同定することができ、実際の治療に役立つ可能性があるほか、下気道における微生物の病態への関与を解析できるツールとなりうることに意義がある。さらに、抗菌薬の感受性についても網羅的に検出することが可能となり、国内における薬剤耐性対策に寄与する可能性があるといえる。

Report

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

    (14 results)

All 2020 2019 2018 2017

All Journal Article (3 results) (of which Peer Reviewed: 3 results,  Open Access: 1 results) Presentation (11 results) (of which Int'l Joint Research: 5 results,  Invited: 2 results)

  • [Journal Article] Rapid and Accurate Species Identification of Mitis Group Streptococci Using the MinION Nanopore Sequencer.2020

    • Author(s)
      Imai K, Nemoto R, Koda M, Tarumoto N, Sakai J, Kawamura T, Ikebuchi K, Mitsutake K, Murakami T, Maesaki S, Fujiwara T, Hayakawa S, Hoshino T, Seki M, Maeda T.
    • Journal Title

      Front Cell Infect Microbiol

      Volume: 11 Pages: 1-11

    • DOI

      10.3389/fcimb.2020.00011

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] An identification protocol for ESBL-producing Gram-negative bacteria bloodstream infections using a MinION nanopore sequencer2019

    • Author(s)
      Sakai Jun、Tarumoto Norihito、Kodana Masahiro、Ashikawa Sae、Imai Kazuo、Kawamura Toru、Ikebuchi Kenji、Murakami Takashi、Mitsutake Kotaro、Maeda Takuya、Maesaki Shigefumi
    • Journal Title

      Journal of Medical Microbiology

      Volume: 68 Issue: 8 Pages: 1219-1226

    • DOI

      10.1099/jmm.0.001024

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Rapid identification of pathogens from positive blood culture bottles with the MinION nanopore sequencer2018

    • Author(s)
      Ashikawa Sae、Tarumoto Norihito、Imai Kazuo、Sakai Jun、Kodana Masahiro、Kawamura Toru、Ikebuchi Kenji、Murakami Takashi、Mitsutake Kotaro、Maesaki Shigefumi、Maeda Takuya
    • Journal Title

      Journal of Medical Microbiology

      Volume: 67 Issue: 11 Pages: 1589-1595

    • DOI

      10.1099/jmm.0.000855

    • Related Report
      2018 Research-status Report
    • Peer Reviewed
  • [Presentation] Constructing the Rapid Detection Methods of Drug-Resistance Mutation of Tuberculosis using mobile nanopore sequencer, MinION2019

    • Author(s)
      Misawa K, Hamamoto T, Imai K, Tarumoto N, Kawana A, Maesaki S, Maeda T
    • Organizer
      29th European Congress of Clinical Microbiology and Infectious Diseases
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research
  • [Presentation] A method using the MinION nanopore sequencer for detecting bacteria and ESBL-producing species in the bloodstream2019

    • Author(s)
      Sakai J, Tarumoto N, Imai K, Maeda T, Maesaki S.
    • Organizer
      29th European Congress of Clinical Microbiology and Infectious Diseases
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research
  • [Presentation] A method using the MinION nanopore sequencer for detecting bacteria and ESBL-producing species in the bloodstream2019

    • Author(s)
      Jun Sakai, Norihito Tarumoto, Kazuo Imai, Takuya Maeda, Shigefumi Maesaki
    • Organizer
      The 29th European Congress of Clinical Microbiology & Infectious Diseases
    • Related Report
      2018 Research-status Report
    • Int'l Joint Research
  • [Presentation] Constructing the rapid detection methods of drug-resistance mutation of tuberculosis using mobile nanopore sequencer MinION2019

    • Author(s)
      Kazuhisa Misawa, Takaaki Hamamoto, Kazuo Imai, Norihito Tarumoto, Akihiko Kawana, Shigefumi Maesaki, Takuya Maeda
    • Organizer
      The 29th European Congress of Clinical Microbiology & Infectious Diseases
    • Related Report
      2018 Research-status Report
    • Int'l Joint Research
  • [Presentation] 血液培養陽性例におけるナノポア型ポータブルシーケンサーMinIONの菌種同定能の検討2019

    • Author(s)
      酒井純、樽本憲人、今井一男、前田卓哉、前崎繁文
    • Organizer
      第93回日本感染症学会 総会
    • Related Report
      2018 Research-status Report
  • [Presentation] ナノポアシーケンサーを使用した結核菌の薬剤耐性に関する遺伝子領域の解析2019

    • Author(s)
      三沢和央、濱本隆明、今井一男、樽本憲人、藤倉雄二、前田卓哉、前崎繁文、川名明彦
    • Organizer
      第93回日本感染症学会 総会
    • Related Report
      2018 Research-status Report
  • [Presentation] Evaluation of MinION sequencer for identification of pathogens from positive blood cultures in bloodstream infections.2018

    • Author(s)
      Takuya Maeda
    • Organizer
      3rd Technology Seminar on the MinION sequencing, Manado, Indonesia.
    • Related Report
      2018 Research-status Report
    • Int'l Joint Research / Invited
  • [Presentation] Practical uses of MinION sequencer at clinical settings in Japan2017

    • Author(s)
      前田卓哉
    • Organizer
      2nd Technology Seminar on the MinION sequencing, Bangkok, Thailand
    • Related Report
      2017 Research-status Report
    • Invited
  • [Presentation] Streptococcus pneumoniaeとStreptococcus mitisの鑑別におけるMinIONの有用性2017

    • Author(s)
      樽本憲人、今井一男、根本莉奈、酒井純、前田卓哉、関みつ子、早川智、村上孝、前﨑繁文
    • Organizer
      第29回臨床微生物学会総会・学術集会
    • Related Report
      2017 Research-status Report
  • [Presentation] MinIONによるストレプトコッカス属鑑別2017

    • Author(s)
      根本莉奈、今井一男、河村 亨、樽本憲人、前田卓哉、村上孝、前﨑繁文
    • Organizer
      第54回日臨技 関甲信支部・首都圏支部医学検査学会
    • Related Report
      2017 Research-status Report
  • [Presentation] 血液培養陽性検体におけるナノポア型シーケンサーの病原微生物同定能の検討2017

    • Author(s)
      蘆川彩瑛、酒井純、河村亨、樽本憲人、前田卓哉、村上孝、前﨑繁文
    • Organizer
      第54回日臨技 関甲信支部・首都圏支部医学検査学会
    • Related Report
      2017 Research-status Report

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

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