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Detection, quantification, and isolation of Escherichia coli O157 by using flow cytometry and fluorescence-activated sorting

Research Project

Project/Area Number 23860003
Research Category

Grant-in-Aid for Research Activity Start-up

Allocation TypeSingle-year Grants
Research Field Civil and environmental engineering
Research InstitutionHokkaido University

Principal Investigator

ISHII Satoshi  北海道大学, 大学院・工学研究院, 助教 (10612674)

Project Period (FY) 2011 – 2012
Project Status Completed (Fiscal Year 2012)
Budget Amount *help
¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000)
Fiscal Year 2012: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2011: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Keywords大腸菌O157 / 水系感染症 / フローサイトメトリ / セルソーティング / FACS / 病原性大腸菌 / 一細胞分離 / 蛍光抗体 / 病原細菌 / 大腸菌0157 / 汚染源特定
Research Abstract

Pathogenic bacteria in irrigation water may attach to vegetables and can cause foodborne diseases. For safe water and food supply, sources of pathogen contamination should be identified and removed. To identify the sources of pathogens, specific detection and isolation of live pathogens is necessary. The objectives of this study were to develop a method to specifically detect and isolate pathogenic bacteria from environmental water samples by using flow cytometry and fluorescence-activated cell sorting (FCM-FACS). We used fluorescence-labeled anti-O157 antibodies to specifically detect E. coli O157 cells. Detected cells were individually sorted and grown in a medium in 96-well plates, and their identities were confirmed as E. coli O157. Even when the proportion of O157 cells in O157/K12 cell mixture was as low as 0.01%, we could specifically detect O157 cells. In addition, we could detect O157 cells present at 10 cells/ml in environmental water samples. Isolated cells can be used to identify their origins by comparing their characteristics to those in known-source E. coli O157 databases.

Report

(3 results)
  • 2012 Annual Research Report   Final Research Report ( PDF )
  • 2011 Annual Research Report
  • Research Products

    (8 results)

All 2013 2012 Other

All Presentation (7 results) Remarks (1 results)

  • [Presentation] 環境水中からのFCM-FACSを用いた大腸菌O157の単離手法の開発2013

    • Author(s)
      小澤就志、石井聡、岡部聡
    • Organizer
      第47回日本水環境学会年会
    • Place of Presentation
      大阪工業大学
    • Related Report
      2012 Final Research Report
  • [Presentation] 環境水中からのFCM-FACSを用いた大腸菌O157の単離手法の開発2013

    • Author(s)
      小澤就志、石井聡、岡部聡
    • Organizer
      第47回 日本水環境学会年会
    • Place of Presentation
      大阪工業大学(大阪市)
    • Related Report
      2012 Annual Research Report
  • [Presentation] Detection, quantification and isolation of Escherichia coli O157 with flow cytometry and fluorescence-activated sorting2012

    • Author(s)
      Ozawa, S., Ishi, S., and Okabe, S
    • Organizer
      Japan and China Joint Symposium, Sustainable Sanitation and Environmentand Human Health Risk Management. Sapporo, Hokkaido, Japan
    • Place of Presentation
      Hokkaido University
    • Year and Date
      2012-10-15
    • Related Report
      2012 Final Research Report
  • [Presentation] Detection of enteropathogenic Escherichia coli O157 with flow cytometry.2012

    • Author(s)
      Ozawa, S., Ishi, S., and Okabe, S.
    • Organizer
      The 2nd Young Professors and Students Symposiumof NSFC-JST Joing Resarch Program.Xi'an, P.R. China.
    • Place of Presentation
      Xi'anUniversity of Architecture and Technology
    • Year and Date
      2012-03-28
    • Related Report
      2012 Final Research Report
  • [Presentation] Detection of enteropathogenic Escherichia coli 0157 with flow cytometry2012

    • Author(s)
      Ozawa, S., Ishi,S., Okabe, S.
    • Organizer
      The 2nd Young Professors and Students Symposium of NSFC-JST Joing Research Program
    • Place of Presentation
      西安建築科学大学(西安、中華人民共和国)
    • Year and Date
      2012-03-28
    • Related Report
      2011 Annual Research Report
  • [Presentation] フローサイトメトリーによる生きた病原性大腸菌O157の特異的検出および分離2012

    • Author(s)
      小澤就志、石井聡、岡部聡
    • Organizer
      第46回日本水環境学会年会
    • Place of Presentation
      東洋大学(東京都)
    • Year and Date
      2012-03-16
    • Related Report
      2011 Annual Research Report
  • [Presentation] フローサイトメ トリーによる生きた病原性大腸菌O157の特異的検出および分離2012

    • Author(s)
      小澤就志、石井聡、岡部聡
    • Organizer
      第46回日本水環境学会年会
    • Place of Presentation
      東洋大学
    • Related Report
      2012 Final Research Report
  • [Remarks]

    • URL

      http://www.eng.hokudai.ac.jp/labo/water/

    • Related Report
      2012 Final Research Report

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Published: 2011-09-05   Modified: 2019-07-29  

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