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

Research Project

Project/Area Number 15390216
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Legal medicine
Research InstitutionFUKUSH1MA 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)
ABE Sumiko  Fukushima Med. Univ., Schl. of Med., Assistant professor, 医学部, 講師 (50136975)
MIZUSAWA Ikubumi  Fukushima Med. Univ., Schl of Med., Research assistant, 医学部, 助手 (40192356)
郡司 啓文  福島県立医科大学, 医学部, 講師 (20234643)
Project Period (FY) 2003 – 2004
Project Status Completed (Fiscal Year 2004)
Budget Amount *help
¥5,600,000 (Direct Cost: ¥5,600,000)
Fiscal Year 2004: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2003: ¥4,200,000 (Direct Cost: ¥4,200,000)
KeywordsDrowning / PCR / Bacteria / Plankton / 常在細菌 / 死後変化
Research Abstract

Neither an acid digestion method nor a PCR method for identifying plankton is useful in cases of death by drowning if inhaled water contains no planktons. So we developed a new PCR method for identifying bacteria and the usefulness of identifying bacterial genes for diagnosing drowning was evaluated.
We examined the bacteria species in blood in the heart and in throat and tracheal swabs obtained from autopsy cases using sterilized instruments. Identification of bacteria in the samples was carried out by the conventional culture method. In throat and trachea swabs obtained from non-drowning cases, Streptococcus (Strep.) and Staphylococcus species were identified, but no bacteria were identified in blood samples. In cases of death by drowning, Strep., Enterobacter and Aeromonas species were identified in throat, trachea and also blood samples. It was thought that the bacteria species that normally localized in the throat and trachea invade into blood through the respiratory tract as well as species localized in water in drowning cases.
Based on these results, we designed primer pairs for membrane protein gene of Strep. salivarius (SL1), which is a common bacteria in the throat, and for ATCC 7966 DNA gyrase subunit B gene of Aero. hydrophila (AH1 and AH2), which has been isolated from various water samples. Both of the cultured strains were obtained from RIKEN BioResource Center. Genomic DNA of both bacteria species was extracted using an AquaPure Genomic DNA kit (BioRad). Employing target DNA as a template, the expected lengths of PCR products amplified with SL1, AH1 and AH2 were 152, 212 and 201 bp, respectively. No PCR product was amplified from human or non-target DNA. With SL1, up to 100 pg of Strep.-DNA was identified, and up to 1 μg of Aero.-DNA was detectable with All or AH2 in each sample mixed with 1 μg of human DNA. The development of this PCR method is a step toward the establishment of a reliable method for diagnosing drowning.

Report

(3 results)
  • 2004 Annual Research Report   Final Research Report Summary
  • 2003 Annual Research Report
  • Research Products

    (8 results)

All 2003 Other

All Journal Article (6 results) Publications (2 results)

  • [Journal Article] Phytoplankton gene detection in drowned rabbits2003

    • Author(s)
      Miwako Suto, et al.
    • Journal Title

      Legal medicine 5・Suppl.1

      Pages: 142-144

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] A Novel PCR Method for Identifying Plankton in Case of Death by Drowning2003

    • Author(s)
      Sumiko Abe, et al.
    • Journal Title

      Medicine, science, and the law 43・1

      Pages: 23-30

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Phytoplankton gene detection in drowned rabbits2003

    • Author(s)
      Miwako Suto, Sumiko Abe, Masahide Nakamura, Toshiyuki Suzuki Toshimitsu Itoh, Hideo Kochi, Gen-ichi Hoshiai, Kouichi Hiraiwa
    • Journal Title

      Legal Medicine. 5

      Pages: 142-144

    • NAID

      10012203254

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] A novel PCR method for identifying plankton in cases of death by drowning2003

    • Author(s)
      Sumiko Abe, Miwako Suto, Masahide Nakamura, Hirobumi Gunji, Toshiyuki Suzuki, Toshimitsu Itoh, Hideo Kochi, Gen-ichi Hoshiai, Kouichi Hiraiwa.
    • Journal Title

      Med., Sci. and Law 43

      Pages: 23-30

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Phytoplankton gene detection in drowned rabbits2003

    • Author(s)
      Miwako Suto, et al.
    • Journal Title

      Legal Medicine 5・Suppl.1

      Pages: 142-144

    • Related Report
      2004 Annual Research Report
  • [Journal Article] A Novel PCR Method for Identfying Plankton in Case of Death by Drowning2003

    • Author(s)
      Sumiko Abe, et al.
    • Journal Title

      Med.Sci.Law 43・1

      Pages: 23-30

    • Related Report
      2004 Annual Research Report
  • [Publications] Miwako Suto: "Phytoplankton gene detection in drowned rabbits"Legal Medicine. 5. 142-144 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Sumiko Abe: "A Novel PCR Method for Identifying Plankton in Case of Death by Drowing"Med.Sci.Law. 43・1. 23-30 (2003)

    • Related Report
      2003 Annual Research Report

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Published: 2003-04-01   Modified: 2016-04-21  

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