Development of new molecular technique (LAMP assay) for determination of death by fresh water drowning

Research Project

Project/Area Number	25460871
Research Category	Grant-in-Aid for Scientific Research (C)
Allocation Type	Multi-year Fund
Section	一般
Research Field	Legal medicine
Research Institution	University of Miyazaki
Principal Investigator	KAKIZAKI Eiji 宮崎大学, 医学部, 准教授 (70284833)
Co-Investigator(Kenkyū-buntansha)	YUKAWA NOBUHIRO 宮崎大学, 医学部, 教授 (30240154) SAKAI MASAHIRO 宮崎大学, 農学部, 教授 (20178536) 小澤周二三重大学, 医学(系)研究科(研究院), 講師 (20379944)
Co-Investigator(Renkei-kenkyūsha)	SONODA AI 宮崎大学, 医学部, 助手 (10762122)
Project Period (FY)	2013-04-01 – 2016-03-31
Project Status	Completed (Fiscal Year 2015)
Budget Amount *help	¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000) Fiscal Year 2015: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000) Fiscal Year 2014: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000) Fiscal Year 2013: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Keywords	法医学 / 溺死 / 水棲微生物 / 水棲細菌 / Aeromonas属 / LAMP法 / プランクトン
Outline of Final Research Achievements	We determined the genus Aeromonas (fresh water bacterioplankton) as microbial marker for diagnosis of drowning in fresh water and tried to develop a new method using LAMP (Loop-Mediated Isothermal Amplification) assay. Present method was specific for standard strains of Aeromonas haydrophila and A. salmonicida. The sensitivity of the method was >10 pg/tube. Moreover, the Aeromons spp. were detectable in blood and/or lung tissues from cadavers retrieved from the rivers (10/11), sea (4/10), near estuaries (5/5) by the new method although they were not detectable in those of non-drowning cadavers (0/14). The Aeromonas spp. were not also detectable in blood and lung tissues from cadavers retrieved from baths (0/6). Thus, this technique has high potential as a novel molecular tool with which to examine whether an immersed victim has drowned.