2026-03-13T21:21:43Z https://miyazaki-u.repo.nii.ac.jp/oai

oai:miyazaki-u.repo.nii.ac.jp:00003879 2024-02-09T07:56:45Z 72 72:54

A new molecular approach to help conclude drowning as a cause of death : Simultaneous detection of eight bacterioplankton species using real-time PCR assays with TaqMan probes 水棲細菌を指標としたMultiplex TaqMan PCR法による溺死の補助診断法 内山, 岳人 ウチヤマ, タケト Uchiyama, Taketo 内山 ウチヤマ Uchiyama 岳人 タケト Taketo open access Drowning, Real-time PCR, TaqMan probe, Bacterioplankton, Diatom test 2013年度 We developed a novel tool for concluding drowning as a cause of death. We designed nine primer pairs to detect representative freshwater or marine bacterioplankton (aquatic bacteria) and then used real-time PCR with TaqMan probes to rapidly and specifically detect them. We previously cultured the genus Aeromonas, which is a representative freshwater bacterial species, in blood samples from 94% of victims who drowned in freshwater and the genera Vibrio and/or Photobacterium that are representative marine bacteria in 88% of victims who drowned in seawater. Based on these results, we simultaneously detected eight species of bacterioplankton (Aeromonas hydrophila, A. salmonicida; Vibrio fischeri, V. harveyi, V. parahaemolyticus; Photobacterium damselae, P. leiognathi, P. phosphoreum) using three sets of triplex real- time PCR assays and TaqMan probes labelled with fluorophores (FAM, NED, Cy5). We assayed 266 specimens (109 blood, 157 tissues) from 43 victims, including 32 who had drowned in rivers, ditches, wells, sea or around estuaries. All lung samples of these 32 victims were TaqMan PCR-positive including the lung periphery into which water does not readily enter postmortem. On the other hand, findings in blood and/or closed organs (kidney or liver) were PCR-positive in 84% of the drowned victims (except for those who drowned in baths) although the conventional test detected diatoms in closed organs in only 44% of the victims. Thus, the results of the PCR assay reinforced those of diatom tests when only a few diatoms were detectable in organs due to the low density of diatoms in the water where they were found. Multiplex TaqMan PCR assays for bacterioplankton were rapid, less laborious and high-throughput as well as sensitive and specific. Therefore, these assays would be useful for routine forensic screening tests to estimate the amount and type of aspirated water. eng doctoral thesis VoR http://hdl.handle.net/10458/4706 https://miyazaki-u.repo.nii.ac.jp/records/3879 http://hdl.handle.net/10458/4641 http://hdl.handle.net/10458/4641 医博甲第414号 博士(医学) 2013-09-20 17601 宮崎大学 https://miyazaki-u.repo.nii.ac.jp/record/3879/files/A414Yhakuron.pdf application/pdf 208.1 kB 2020-06-21 https://miyazaki-u.repo.nii.ac.jp/record/3879/files/A414Yshinsa.pdf application/pdf 82.5 kB 2020-06-21 https://miyazaki-u.repo.nii.ac.jp/record/3879/files/A414Honbun.pdf application/pdf 2.4 MB 2020-06-21