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Development of simple, rapid and low-cost onsite simultaneous measurement technique of indicator bacteria based on fluorescence probes

Research Project

Project/Area Number 19K21979
Research Category

Grant-in-Aid for Challenging Research (Exploratory)

Allocation TypeMulti-year Fund
Review Section Medium-sized Section 22:Civil engineering and related fields
Research InstitutionHokkaido University

Principal Investigator

Satoh Hisashi  北海道大学, 工学研究院, 教授 (80326636)

Co-Investigator(Kenkyū-buntansha) 羽深 昭  北海道大学, 工学研究院, 助教 (30735353)
齋藤 伸吾  埼玉大学, 理工学研究科, 教授 (60343018)
Project Period (FY) 2019-06-28 – 2021-03-31
Project Status Completed (Fiscal Year 2020)
Budget Amount *help
¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000)
Fiscal Year 2020: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
Fiscal Year 2019: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
Keywordsセンサ / 大腸菌 / センサー / 計測技術
Outline of Research at the Start

本研究では蛍光プローブを用いて、極めて簡易に、迅速に、低コストで、オンサイトで国内外の環境基準として設定されている複数の指標細菌数を一斉に計測できる技術を開発する。細菌の測定が記憶に残るのは、作業に多大な労力を要するためであろう。本法が完成すれば極めて簡易に細菌を定量できるようになり、特に熟練の技術者がおらず分析にコストをかけられない開発途上国の人の健康と環境を守ることができるようになる。さらに現在は指標細菌数を測定するまでに24時間が必要であるが、本法ではわずか2時間で測定が終了するので、指標細菌数が基準値を超えていた場合に迅速に対応することが可能となる。

Outline of Final Research Achievements

We developed a simple, rapid, and low-cost simultaneous on-site indicator bacteria measurement technique using fluorescent probes. A sample with 40 cfu/L of E. coli could be detected in about 7 hours, and a sample with 4 cfu/L in about 13 hours. The operation is extremely simple: even with untreated sewage, the sample water is placed in a liquid medium or agar medium without any pretreatment, and the fluorescence intensity is automatically measured once every 10 minutes using a microplate reader set at 37°C. Since the number of coliform bacteria can be measured in about three hours for normal treated sewage water, the number of coliform bacteria can be analyzed with high frequency in both time and space, and deterioration of treatment efficiency can be quickly identified. Since the measurement medium is about 0.02 mL, the cost per sample is about 2 Japanese yen.

Academic Significance and Societal Importance of the Research Achievements

極めて簡易にオンサイトで大腸菌を計測できるようになり、特に熟練の技術者がおらず分析にコストをかけられない開発途上国の人の健康と環境を守ることができる。現在は指標細菌数を計測するまでに約24時間かかるが、本法ではわずか3時間で計測が終了するので、指標細菌数が基準値を超えた場合に迅速に対応することが可能となる。現在世界中で広く用いられている計測キットのコストは1サンプル当たり約1300円であるが、本法は1サンプル2円であるので、開発途上国でも問題なく使用できる。一度に96サンプル測定できるので高頻度で指標細菌の経時変化を追う、広範囲の指標細菌の分布を計測する、などが可能となる。

Report

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

    (8 results)

All 2021 2020 2019 Other

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

  • [Int'l Joint Research] University of Ruhuna/University of Peradeniya(スリランカ)

    • Related Report
      2019 Research-status Report
  • [Journal Article] Simple enumeration of Escherichia coli concentrations in river water samples by measuring β-d-glucuronidase activities in a microplate reader2021

    • Author(s)
      Satoh Hisashi、Katayose Yutaka、Hirano Reiko
    • Journal Title

      Water Science and Technology

      Volume: 83 Issue: 6 Pages: 1399-1406

    • DOI

      10.2166/wst.2021.072

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Simple and reliable enumeration of Escherichia coli concentrations in wastewater samples by measuring β-d-glucuronidase (GUS) activities via a microplate reader2020

    • Author(s)
      Satoh Hisashi、Kikuchi Kai、Katayose Yutaka、Tsuda Shu、Hirano Reiko、Hirakata Yuga、Kitajima Masaaki、Ishii Satoshi、Oshiki Mamoru、Hatamoto Masashi、Takahashi Masahiro、Okabe Satoshi
    • Journal Title

      Science of The Total Environment

      Volume: 715 Pages: 136928-136928

    • DOI

      10.1016/j.scitotenv.2020.136928

    • Related Report
      2019 Research-status Report
    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] Development of a simple and rapid method for determining coliforms in wastewater based on fluorogenic enzymatic substrate2019

    • Author(s)
      佐藤 久、津田 収、菊地 凱、平野 麗子
    • Journal Title

      Journal of Japan Sewage Works Association

      Volume: 56 Issue: 684 Pages: 110-117

    • DOI

      10.24748/jswa.56.684_110

    • NAID

      130007722331

    • ISSN
      0021-4639, 2434-2475
    • Year and Date
      2019-10-01
    • Related Report
      2019 Research-status Report
    • Peer Reviewed / Open Access
  • [Presentation] 簡易大腸菌分析法による下水中大腸菌数の測定2020

    • Author(s)
      深田 翔介・佐藤 久
    • Organizer
      第54回日本水環境学会年会
    • Related Report
      2019 Research-status Report
  • [Presentation] A Simple and Rapid Method for Enumerating Escherichia coli in Wastewater by Measuring β-D-glucuronidase Activity2019

    • Author(s)
      H. Satoh, K. Kikuchi, Y. Katayose, S. Tsuda, R. Hirano, S. Ishii, S. Okabe
    • Organizer
      IWA Water and Development Congress & Exhibition 2019
    • Related Report
      2019 Research-status Report
    • Int'l Joint Research
  • [Remarks] 佐藤久研究グループWebsite

    • URL

      https://www.eng.hokudai.ac.jp/labo/aqua/contents/HisashiSatoh/index-HisashiSatoh.html

    • Related Report
      2020 Annual Research Report
  • [Remarks] 北大 佐藤久研究グループ

    • URL

      https://www.eng.hokudai.ac.jp/labo/aqua/contents/HisashiSatoh/index-HisashiSatoh.html

    • Related Report
      2019 Research-status Report

URL: 

Published: 2019-07-04   Modified: 2022-01-27  

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