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Simultaneous detection of multiple aquatic microbes using TaqMan PCR system

Research Project

Project/Area Number 17K09268
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Research Field Legal medicine
Research InstitutionUniversity of Miyazaki

Principal Investigator

Yukawa Nobuhiro  宮崎大学, 医学部, 教授 (30240154)

Co-Investigator(Kenkyū-buntansha) 柿崎 英二  宮崎大学, 医学部, 准教授 (70284833)
園田 愛  宮崎大学, 医学部, 助手 (10762122)
Project Period (FY) 2017-04-01 – 2020-03-31
Project Status Completed (Fiscal Year 2019)
Budget Amount *help
¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
Fiscal Year 2019: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2018: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2017: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
Keywords溺死の診断 / リアルタイムPCR / 水棲微生物 / 珪藻 / 水棲細菌 / 法医学
Outline of Final Research Achievements

We developed a new real-time PCR method to detect fresh and marine diatoms for diagnosis of drowning in various water environments. Present method was specific for various standard strains of fresh and marine diatoms (Achnantidium, Aulacoseira, Nitzschia, Tabellaria, Ulnaria, etc. as fresh water diatoms; Eucampia, Odontella, Pseudo-nitzschia, Thalassionema, etc. as marine diatoms). The sensitivity of the method was >10 pg/tube. Moreover, the diatom DNAs were detectable in water samples collected from various water environments although they were undetectable in the blood and tissues (kidney and liver) from cadavers retrieved from the rivers and sea by the new method. These results gave us one of the reasons to prove our hypothesis that the diatom DNAs would be almost undetectable in blood and closed organs (kidney and liver).

Academic Significance and Societal Importance of the Research Achievements

海水域や淡水域など生息水域に限らず,一つのプライマーセットを用いて一括して珪藻類を検出可能な測定系の開発に取り組んだ。開発した測定系は珪藻以外の他の水棲微生物との交叉性(偽陽性)を示さず,検出限界は1~10 pg/tubと良好であった。ところで従来の珪藻検査から得られる結果の意義,特に肺以外の諸臓器や血液から得られる結果の意義については,世界中で意見が分かれ未だ解決には至っていない。本研究によって得られた成果はこの問題に対しいくつかの重要な示唆を与えることができたと考えている。今後も引き続き検討を重ねこの課題の解決に繋げたい。

Report

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

    (18 results)

All 2019 2018 2017

All Journal Article (7 results) (of which Peer Reviewed: 6 results) Presentation (11 results) (of which Int'l Joint Research: 2 results)

  • [Journal Article] Acceleration effect of the forensic luminol reaction induced by visible light irradiation of whole human blood aqueous solutions2019

    • Author(s)
      Hayashi S, Kakizaki E, Sonoda A, Shinkawa N, Shiragami T*, Yukawa N*
    • Journal Title

      Forensic Science International

      Volume: 299 Pages: 208-214

    • DOI

      10.1016/j.forsciint.2019.04.007

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Neonatal Death Caused by Interrupted Aortic Arch Associated With 22q11.2 Deletion Syndrome: An Autopsy Case Report2019

    • Author(s)
      Shinkawa N*, Yamaguchi M, Ozaki M, Yukawa N
    • Journal Title

      American Journal of Forensic Medicine and Pathology

      Volume: 40 Issue: 2 Pages: 178-182

    • DOI

      10.1097/paf.0000000000000454

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Simple detection of bacterioplankton using a loop-mediated isothermal amplification (LAMP) assay: First practical approach to 72 cases of suspected drowning2018

    • Author(s)
      Kakizaki E, Sonoda A, Sakai M and Yukawa N
    • Journal Title

      Forensic Science International

      Volume: 289 Pages: 289-303

    • DOI

      10.1016/j.forsciint.2019.02.008

    • Related Report
      2019 Annual Research Report 2018 Research-status Report
    • Peer Reviewed
  • [Journal Article] 小児のボタン電池誤飲を防ぐために - 低電圧1.5Vのボタン電池でも組織傷害性アルカリは生成される2018

    • Author(s)
      新川慶明,中野 敦,柿崎英二,林 里采,園田 愛,湯川修弘
    • Journal Title

      日本医事新報

      Volume: 4896 Pages: 20-22

    • NAID

      40021472235

    • Related Report
      2018 Research-status Report
    • Peer Reviewed
  • [Journal Article] 覚せい剤(メタンフェタミン)の法医学講義ノート2018

    • Author(s)
      林 里采,石田 康,園田 愛,新川慶明,柿崎英二,湯川修弘
    • Journal Title

      法医学の実際と研究

      Volume: 61 Pages: 201-216

    • Related Report
      2018 Research-status Report
    • Peer Reviewed
  • [Journal Article] 特集 次世代シーケンサーが可能にした感染学の新しい展開,4. 臨床への応用(Clinical Sequencing),4)法医学への応用2017

    • Author(s)
      柿崎英二,湯川修弘
    • Journal Title

      化学療法の領域

      Volume: 33 Pages: 143-151

    • Related Report
      2017 Research-status Report
  • [Journal Article] 一酸化炭素中毒の法医学講義ノート(Haldaneの第一法則・第二法則)2017

    • Author(s)
      湯川修弘,園田 愛,柿崎英二,落合秀信
    • Journal Title

      法医学の実際と研究

      Volume: 60 Pages: 219-235

    • Related Report
      2017 Research-status Report
    • Peer Reviewed
  • [Presentation] 水棲微生物を指標とした溺死の補助診断法:最も少ない労力で検査を行うためのプロトコールの最適化2019

    • Author(s)
      園田 愛,柿崎英二,新川慶明,湯川修弘
    • Organizer
      第103次日本法医学会学術全国集会(仙台)
    • Related Report
      2019 Annual Research Report
  • [Presentation] 慢性硬膜下出血内に認められたヘマトイジン含有マクロファージと思われる黄色色素含有細胞2019

    • Author(s)
      新川慶明,園田 愛,松山美紀,柿崎英二,林 里采,和田 啓,湯川修弘
    • Organizer
      第103次日本法医学会学術全国集会(仙台)
    • Related Report
      2019 Annual Research Report
  • [Presentation] 血液水溶液への可視光照射によるルミノール反応の加速効果2019

    • Author(s)
      林 里采,白上 努,柿崎英二,新川慶明,園田 愛,湯川修弘
    • Organizer
      第69回日本法医学会学術九州地方集会(大分)
    • Related Report
      2019 Annual Research Report
  • [Presentation] 切断された遺体に骨化性筋炎を認め,生前の暴行が疑われた一例2019

    • Author(s)
      新川慶明,柿崎英二,林 里采,園田 愛,湯川修弘
    • Organizer
      第69回日本法医学会学術九州地方集会(大分)
    • Related Report
      2019 Annual Research Report
  • [Presentation] A new enzymatic method for extracting diatoms from lung, kidney and liver tissues of suspected drowning cases using papain digestion: First practical application2018

    • Author(s)
      Kakizaki E, Sonoda A, Shinkawa N, Yukawa N
    • Organizer
      24th Congress of the International Academy of Legal Medicine (IALM)
    • Related Report
      2018 Research-status Report
    • Int'l Joint Research
  • [Presentation] Autopsy of a body with discolored sun-exposed areas of skin, and a literature survey on postmortem suntan2018

    • Author(s)
      Yukawa N, Shinkawa N, Sonoda A, Kakizaki E, Hayashi S
    • Organizer
      24th Congress of the International Academy of Legal Medicine (IALM)
    • Related Report
      2018 Research-status Report
    • Int'l Joint Research
  • [Presentation] 右手に笹の葉を硬く握った状態で発見され即時性死体硬直と思われた溺死の一解剖例2018

    • Author(s)
      湯川修弘,新川慶明,柿崎英二,園田 愛,林 里采,小片 守
    • Organizer
      第68回日本法医学会学術九州地方集会
    • Related Report
      2018 Research-status Report
  • [Presentation] ボタン電池の誤飲や嵌頓において,低電圧1.5 Vの電池でも組織傷害性のアルカリが生成される理由について2018

    • Author(s)
      新川慶明,柿崎英二,林 里采,園田 愛,湯川修弘
    • Organizer
      第68回日本法医学会学術九州地方集会
    • Related Report
      2018 Research-status Report
  • [Presentation] 水中死体の肺及び血中に存在する水棲微生物の網羅的解析(第2報):法医解剖32例のメタ16S解析2017

    • Author(s)
      柿崎英二,小椋 義俊,林 哲也,湯川修弘
    • Organizer
      第101次日本法医学会学術全国集会(岐阜)
    • Related Report
      2017 Research-status Report
  • [Presentation] 溺死診断のためのLAMP法を用いた簡易検査法の開発(第2報):Vibrio属・Photobacterium属細菌の一括検出2017

    • Author(s)
      園田 愛,柿崎英二,酒井正博,湯川修弘
    • Organizer
      第101次日本法医学会学術全国集会(岐阜)
    • Related Report
      2017 Research-status Report
  • [Presentation] 一酸化炭素中毒の法医学講義ノート(Haldaneの第一法則・第二法則)2017

    • Author(s)
      湯川修弘,園田 愛,柿崎英二,林 里采
    • Organizer
      第67回日本法医学会学術九州地方集会(那覇)
    • Related Report
      2017 Research-status Report

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

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