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A new enzymatic method for extracting diatoms from organs of suspected drowning cases using papain digestion

Research Project

Project/Area Number 18K10132
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Review Section Basic Section 58040:Forensics medicine-related
Research InstitutionUniversity of Miyazaki

Principal Investigator

Kakizaki Eiji  宮崎大学, 医学部, 准教授 (70284833)

Co-Investigator(Kenkyū-buntansha) 園田 愛  宮崎大学, 医学部, 助手 (10762122)
新川 慶明  宮崎大学, 医学部, 助教 (40625836)
湯川 修弘  宮崎大学, 医学部, 教授 (30240154)
Project Period (FY) 2018-04-01 – 2021-03-31
Project Status Completed (Fiscal Year 2020)
Budget Amount *help
¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2020: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2019: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2018: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Keywords法医学 / 溺死の診断 / プランクトン検査 / パパイン(Papain) / タンパク質分解酵素 / 珪藻 / プロティナーゼK(Proteinase K) / Papain / 蛋白質分解酵素 / Proteinase K
Outline of Final Research Achievements

Diatom analysis is very effective for positive diagnosis of water inhalation in drowning. However, conventional strong acid diatom testing is laborious and potentially dangerous. We propose a simple, fast, and safe protocol using inexpensive reagents such as papain, SDS, and 5 N HCl for extracting diatoms from lung, kidney, and liver tissues. First, we determined optimal conditions for papain digestion using porcine tissues. Papain digestion was clearly superior to Proteinase K digestion. Next, for assessing the assay effectiveness in practical cases, the papain digestion protocol was applied to 80 tissue samples from 20 suspected drowning victims. The proposed method can be useful as a less-laborious, less-hazardous, and less-costly minimal test.

Academic Significance and Societal Importance of the Research Achievements

すべての水中死体に対して,時間と労力のかかる従来の検査法(壊機法)を行うことは簡単ではない。またこの検査は加熱した強酸(発煙硝酸)を用いるため,安全面(化学熱傷,酸性ガスの吸引)や設備面(遠心機及び周辺機器への金属腐食,ドラフトチャンバーの設置・維持管理)の問題も抱えている。そこで本研究では植物由来のタンパク質分解酵素Papainを用いた新しい検査法を開発し,従来の壊機法によるプランクトン検査の代替法として利用できることを明らかにした。このように簡便・迅速かつ安全・安価な珪藻検査の方法は国内外を問わず報告例はなく,今後新しい検査法として死因究明や犯罪捜査に貢献できると考えている。

Report

(4 results)
  • 2020 Annual Research Report   Final Research Report ( PDF )
  • 2019 Research-status Report
  • 2018 Research-status Report
  • Research Products

    (20 results)

All 2021 2020 2019 2018

All Journal Article (9 results) (of which Peer Reviewed: 8 results) Presentation (11 results) (of which Int'l Joint Research: 2 results)

  • [Journal Article] Adult dismembered body with myositis ossificans: Evidence for physical abuse in an autopsy case report2021

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

      American Journal of Forensic Medicine and Pathology

      Volume: 42 Issue: 1 Pages: 73-76

    • DOI

      10.1097/paf.0000000000000628

    • Related Report
      2020 Annual Research 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(第2報)2020

    • 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: 5040 Pages: 48-50

    • NAID

      40022420735

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed
  • [Journal Article] 亜硝酸ナトリウムとアスコルビン酸/塩の混合中毒?【レター】2020

    • Author(s)
      新川慶明*,湯川修弘
    • Journal Title

      法医学の実際と研究

      Volume: 63 Pages: 178-178

    • Related Report
      2020 Annual Research Report
  • [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 Research-status 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 Research-status 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 Research-status 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.2018.05.035

    • Related 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
  • [Presentation] 壊機法によるプランクトン検査:偽陽性を生じる要因の一つについて2020

    • Author(s)
      柿崎英二,園田 愛,柳井章江,新川慶明,湯川修弘
    • Organizer
      第104次日本法医学会学術全国集会(京都)
    • Related Report
      2020 Annual Research Report
  • [Presentation] Evaluation of photochemical damage of visible-light-irradiated blood by forensic luminol reaction2020

    • Author(s)
      林 里采,白上 努,松本 仁,柿崎英二,新川慶明,園田 愛,湯川修弘
    • Organizer
      第104次日本法医学会学術全国集会(京都)
    • Related Report
      2020 Annual Research Report
  • [Presentation] Degree of reductionを用いたアルコール代謝の法医学教育2020

    • Author(s)
      湯川修弘,新川慶明,林 里采,園田 愛,柿崎英二
    • Organizer
      第70回日本法医学会学術九州地方集会(福岡)
    • Related Report
      2020 Annual Research Report
  • [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 Research-status 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 Research-status 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 Research-status 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 Research-status 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

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Published: 2018-04-23   Modified: 2022-01-27  

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