A new enzymatic method for extracting diatoms from organs of suspected drowning cases using papain digestion
| Project/Area Number |
18K10132
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Review Section |
Basic Section 58040:Forensics medicine-related
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| Research Institution | University of Miyazaki |
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Principal Investigator |
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| 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)
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| Project Period (FY) |
2018-04-01 – 2021-03-31
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| Project Status |
Completed (Fiscal Year 2020)
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| 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)
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| 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 |
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| 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.
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| 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。
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Report
(4 results)
Research Products
(20 results)
