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A novel PCR method for identifying plankton in cases of death by drowning

Research Project

Project/Area Number 13670430
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Legal medicine
Research InstitutionFUKUSHIMA MEDICAL UNIVERSITY

Principal Investigator

HIRAIWA Kouichi  Fukushima Med. Univ., Schl. Of Med., Professor, 医学部, 教授 (60124616)

Co-Investigator(Kenkyū-buntansha) KURISAKI Emiko  Fukushima Med. Univ., Schl. Of Med., Assistant Professor, 医学部, 講師 (30106356)
GUNJI Hirobumi  Fukushima Med. Univ., Schl. Of Med., Assistant Professor, 医学部, 講師 (20234643)
ABE Sumiko  Fukushima Med. Univ., Schl. Of Med., Research Assistant, 医学部, 助手 (50136975)
MIZUSAWA Ikumumi  Fukushima Med. Univ., Schl. Of Med., Research Assistant, 医学部, 助手 (40192356)
星合 愿一  宮城県水産研究開発センター, 部長(研究職)
Project Period (FY) 2001 – 2002
Project Status Completed (Fiscal Year 2002)
Budget Amount *help
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 2002: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 2001: ¥2,300,000 (Direct Cost: ¥2,300,000)
KeywordsPlankton / PCR / Chlorophyll-related sequences / drowning
Research Abstract

We developed a new PCR method for identifying plankton in cases of death by drowning. We designed 4 primer pairs for chlorophyll-related genes of Euglena gracilis (EG) and Skeletonema costatum (SK), which are commonly distributed in water. The primers were selected from sequences coding chloroplast/chlorophyll apoprotein of EG (EG1 and EG2) and fucoxanthin-chlorophyll a/c harvesting protein of SK (SK1 and SK2). With EG1 or EG2, up to 2 fg of EG-DNA was identified, and 0.2 pg of SK-DNA was detectable with SK1 or SK2. No PCR products were amplified from green vegetables (komatsuna, spinach, parsley) or human DNA with the 4 primer pairs. Regardless of origin, seawater or fresh water, most diatoms were detectable with primer pairs of EG1 and EG2. With SK1, only Centrales diatoms were identified, and 5 diatom strains originating from seawater were detectable with SK2. EG1 and EG2 gave rise to PCR products from most water samples. By using Percoll, plankton was easily isolated from human tissue and blood samples with which various water samples obtained from the sea, a river and a dam had been mixed. Plankton genes were detected in lung, liver and kidney homogenates mixed with 1 to 3 ml of water samples. By using Percoll, plankton was easily isolated from human tissue or blood samples, and good results of PCR analysis were obtained in cases of death by drowning. PCR products were amplified from lung, liver and kidney samples obtained from drowning victims that showed diatom-negative results using the acid digestion method. In order to evaluate the usefulness of the plankton method for diagnosis of drowning, we examined plankton genes in a non-drowned rabbit submerged after death. Plankton genes were detected in lung samples but not in heart blood samples. Because of postmortem plankton invasion into lungs, lungs are not suitable for plankton identification.

Report

(3 results)
  • 2002 Annual Research Report   Final Research Report Summary
  • 2001 Annual Research Report
  • Research Products

    (17 results)

All Other

All Publications (17 results)

  • [Publications] 須藤美和子: "PCR法による植物プランクトンの検出"法医学の実際と研究. 44. 79-84 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 須藤美和子: "PCR法による植物プランクトンの検出-第3報-"日本法医学雑誌. 55・1. 88 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 須藤美和子: "実験動物を用いた植物プランクトンの検出"日本法医学雑誌. 56・1. 103 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Miwako Suto: "PCR detection of phytoplankton genes in a case of death by drowning"Fifth International Symposium on ADVANCES IN LEAGAL MEDICINE Abstracts. 65 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 須藤 美和子: "PCR法によるヒト組織からの植物プランクトンの検出"法医学の実際と研究. 45. 93-98 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Sumiko Abe: "A novel PCR method for identifying plankton in cases of death by drowning"Medicine, Science and Law. 23-30 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Miwako Suto, Sumiko Abe, Masahide Nakamura, Toshiyuki Suzuki, Toshimitsu Itoh, Hideo Kochi, Kouichi hiraiwa: "Detection of phytoplankton genes by PCR"The Research and Practice in Forensic Medicine. 44. 79-84 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Miwako Suto, Sumiko Abe, Masahide Nakamura, Toshiyuki Suzuki, Toshimitsu Itoh, Hideo Kochi, Gen-ichi Hoshiai, Kouichi Hiraiwa: "PCR detection of phytoplankton genes in a case of death by drowning"Fifth International symposium on ADVANCES IN LEGAL MEDICINE Abstract. 65. (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Miwako Suto, Sumiko Abe, Masahide Nakamura, Toshiyuki Suzuki, Toshimitsu Itoh, Hideo Kochi, Gen-ichi Hoshiai, Kouichi Hiraiwa: "Detection of phytoplankton genes by PCR from human tissue"The Research and Practice in Forensic Medicine. 45. 93-98 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Sumiko Abe, Miwako Suto, Masahide Nakamura, Toshiyuki Suzuki, Toshimitsu Itoh, Hideo Kochi, Gen-ichi Hoshiai, Kouichi Hiraiwa: "A novel PCR method for identifying plankton in cases of death by drowning"Med., Sci. and Law. 43. 23-30 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Miwako Suto: "PCR detection of phytoplankton genes in a case of death by drowning"Fifth International Symposium on ADVANCES IN LEAGAL MEDICINE Abstracts. 65 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] 須藤 美和子: "PCR法によるヒト組織からの植物プランクトンの検出"法医学の実際と研究. 45. 93-98 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Sumiko Abe: "A novel PCR method for identifying plankton in cases of death by drowning"MEDICINE, SCIENCE and LAW. (in print). (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] 須藤美和子: "PCR法による植物プランクトンの検出-第2報-"法医学の実際と研究. 44. 326-326 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] 須藤美和子: "PCR法による植物プランクトンの検出"法医学の実際と研究. 44. 79-84 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] 須藤美和子: "PCR法による植物プランクトンの検出-第3報-"日本法医学雑誌. 55・1. 88-88 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] 須藤美和子: "実験動物を用いた植物プランクトンの検出"日本法医学雑誌. 56・1. 103-103 (2002)

    • Related Report
      2001 Annual Research Report

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Published: 2001-04-01   Modified: 2016-04-21  

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