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Development of a rapid and easy genotyping method for identification and monitoring transmission events by hypervirulent Klebsiella pneumoniae

Research Project

Project/Area Number 20K08819
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Review Section Basic Section 54030:Infectious disease medicine-related
Research InstitutionNagoya University

Principal Investigator

Kawamura Kumiko  名古屋大学, 医学系研究科(保健), 准教授 (30335054)

Co-Investigator(Kenkyū-buntansha) 八木 哲也  名古屋大学, 医学系研究科, 教授 (70333573)
鈴木 匡弘  藤田医科大学, 医学部, 准教授 (70446649)
Project Period (FY) 2020-04-01 – 2023-03-31
Project Status Completed (Fiscal Year 2022)
Budget Amount *help
¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2022: ¥650,000 (Direct Cost: ¥500,000、Indirect Cost: ¥150,000)
Fiscal Year 2021: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2020: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
Keywords高病原性肺炎桿菌 / カルバペネマーゼ産生腸内細菌目細菌 / multiplex PCR法 / 菌種マーカー / sequence type / 莢膜型K1 / 病原遺伝子(rmpA/A2) / 薬剤耐性遺伝子 / 高病原性肺炎桿菌(hvKp) / カルバペネマーゼ産生腸内細菌目細菌(CPE) / sequence type (ST) / 病原遺伝子 (rmpA/A2) / 高病原性肺炎桿菌 (hvKp) / カルバペネマーゼ産生腸内細菌科細菌 (CPE) / multiplex PCR 法 / 肺炎桿菌 / 高病原性 / 感染症診断 / 感染制御 / 解析法の構築
Outline of Research at the Start

高病原性肺炎桿菌(hvKp)の出現と増加、さらにはカルバペネマーゼを産生するhvKp株(CPE-hvKp)の出現も報告され、肺炎桿菌感染症の現状は深刻になりつつある。hvKpおよびCPE-hvKp感染症は市中および医療環境の両方でみられることから、本感染症の診断と感染制御の両方に貢献できる分子疫学解析法が必要である。

本研究では、CPEの世界的流行クローンや病原性と関連するシークエンスタイプの特定、hvKp株に特異的な病原遺伝子群の保有や莢膜型別ならびにカルバペネマーゼ産生遺伝子の保有をOne-stepで検出する "multiplex PCR typing method" を構築する。

Outline of Final Research Achievements

We aimed to develop the multiplex PCR typing method for K. pneumoniae that would be useful in hospital epidemiology and outbreak investigation. This typing method consists of two multiplex PCR reactions that were designed to detect 25 ORFs specific to bacterial genetic lineages, species, antimicrobial-resistant genes (blaCTX-M, blaIMP and blaKPC), a capsular K1-specific gene and a virulence factor gene (rmpA/A2). The distribution patterns of ORFs among K. pneumoniae correlated well with multilocus sequence typing, and closely related species could be distinguished and antimicrobial resistance and hypervirulence genes were identified. This typing method is a rapid and easy genotyping method for monitoring transmission events by K. pneumoniae in clinical microbiology laboratories, and supplies clear and informative molecular typing results for the isolates. In addition, this would facilitate molecular epidemiological analysis in infection control and hospital epidemiology investigations.

Academic Significance and Societal Importance of the Research Achievements

本研究で構築したmultiplex PCR typing methodは、病原性やアウトブレイクと関連する遺伝子型や莢膜型(K1)、薬剤耐性遺伝子および病原遺伝子の保有をOne-stepで解析する分子疫学解析法であり、分子疫学解析標準法と同等の性能を有していた。日常検査における本typing methodの導入は、肺炎桿菌とその近縁種の分布、カルバペネム耐性株および高病原性株の流布の実態を明らかにするとともに、日常診療におけるそれら菌株識別の有用性を明らかにすることに貢献するものと考える。加えて、それらの結果は病院の感染制御および院内感染への対策強化にも貢献することが期待される。

Report

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

    (10 results)

All 2023 2022 2021 2020

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

  • [Journal Article] Molecular Epidemiology of <i>Enterobacter cloacae</i> Complex Isolates with Reduced Carbapenem Susceptibility Recovered by Blood Culture2022

    • Author(s)
      Sarangi Jayathilake, Matsuo Nao, Nonogaki Rina, Hayashi Michiko, Kawamura Kumiko, Suzuki Masahiro, Jin Wanchun, Tamai Kiyoko, Ogawa Miho, Wachino Jun-ichi, Kimura Kouji, Yagi Tetsuya, Arakawa Yoshichika
    • Journal Title

      Japanese Journal of Infectious Diseases

      Volume: 75 Issue: 1 Pages: 41-48

    • DOI

      10.7883/yoken.JJID.2021.141

    • NAID

      130008143965

    • ISSN
      1344-6304, 1884-2836
    • Year and Date
      2022-01-31
    • Related Report
      2021 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Dissecting the clonality of I1 plasmids using ORF-based binarized structure network analysis of plasmids (OSNAp)2022

    • Author(s)
      Suzuki Masahiro、Norizuki Chihiro、Wachino Jun-ichi、Kawamura Kumiko、Nagano Noriyuki、Nagano Yukiko、Hayashi Wataru、Kimura Kouji、Doi Yohei、Arakawa Yoshichika
    • Journal Title

      Journal of Infection and Chemotherapy

      Volume: 28 Issue: 4 Pages: 473-479

    • DOI

      10.1016/j.jiac.2021.12.003

    • Related Report
      2022 Annual Research Report 2021 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] PCR-based ORF typing of Klebsiella pneumoniae for rapid identification of global clones and transmission events.2022

    • Author(s)
      Nonogaki R, Iijima A, Kawamura K, Kayama S, Sugai M, Yagi T, Arakawa Y, Doi Y, Suzuki M.
    • Journal Title

      J Appl Microbiol

      Volume: 133 Issue: 3 Pages: 2050-2062

    • DOI

      10.1111/jam.15701

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Practical Agar-Based Disk Diffusion Tests Using Sulfamoyl Heteroarylcarboxylic Acids for Identification of Subclass B1 Metallo-β-Lactamase-Producing <i>Enterobacterales</i>2021

    • Author(s)
      Norizuki Chihiro、Wachino Jun-ichi、Jin Wanchun、Kimura Kouji、Kawamura Kumiko、Nagano Noriyuki、Arakawa Yoshichika
    • Journal Title

      Journal of Clinical Microbiology

      Volume: 59 Issue: 10

    • DOI

      10.1128/jcm.00761-21

    • Related Report
      2021 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Characterization of blaCTX-M-27/F1:A2:B20 Plasmids Harbored by Escherichia coli ST131 Sublineage C1/H30R Isolates Spreading among Elderly Japanese in Non-acute Care Settings2020

    • Author(s)
      Matsuo Nao、Nonogaki Rina、Hayashi Michiko、Wachino Jun-ichi、Suzuki Masahiro、Arakawa Yoshichika、Kawamura Kumiko
    • Journal Title

      Antimicrobial Agents and Chemotherapy

      Volume: 64 Issue: 5

    • DOI

      10.1128/aac.00202-20

    • Related Report
      2020 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Dissemination of IncF group F1:A2:B20 plasmid-harbouring multidrug-resistant Escherichia coli ST131 before the acquisition of bla CTX-M in Japan2020

    • Author(s)
      Michiko Hayashi, Mari Matsui, Tsuyoshi Sekizuka, Ayaka Shima, Takaya Segawa, Makoto Kuroda, Kumiko Kawamura, Satowa Suzuki
    • Journal Title

      J Glob Antimicrob Resist

      Volume: 23 Pages: 456-465

    • DOI

      10.1016/j.jgar.2020.10.021

    • Related Report
      2020 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Fatal fulminant community-acquired pneumonia caused by hypervirulent Klebsiella pneumoniae K2-ST862020

    • Author(s)
      Hiroyuki Yamamoto, Anna Iijima, Kumiko Kawamura, Yasuo Matsuzawa, Masahiro Suzuki, Yoshichika Arakawa
    • Journal Title

      Medicine

      Volume: 99 Issue: 21 Pages: e20360-e20360

    • DOI

      10.1097/md.0000000000020360

    • Related Report
      2020 Research-status Report
    • Peer Reviewed / Open Access
  • [Presentation] 敗血症・血流感染患者由来Klebsiella pneumoniaeおよび近縁2菌種(K.variicola, K.quasipneumoniae)の分子疫学的特徴の解析2023

    • Author(s)
      磯貝美結、鈴木匡弘、川村久美子
    • Organizer
      2022年度東海乳酸菌研究会
    • Related Report
      2022 Annual Research Report
  • [Presentation] Practical agar-based disk-diffusion tests for the identification of subclass B1 metallo-β-lactamase-producing Enterobacterales using sulfamoyl heteroarylcarboxylic acids2022

    • Author(s)
      法月千尋、和知野純一、Jin Wanchun, 木村幸司、川村久美子、荒川宜親
    • Organizer
      第33回日本臨床微生物学会総会・学術集会
    • Related Report
      2022 Annual Research Report 2021 Research-status Report
  • [Presentation] 感染症診断と感染制御支援のための新規高病原性肺炎桿菌解析法の構築とその評価 -分子疫学解析手法としてのPOT法の構築2022

    • Author(s)
      野々垣里奈、鈴木匡弘、川村久美子
    • Organizer
      2021年度東海乳酸菌研究会
    • Related Report
      2022 Annual Research Report

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

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