Uchigasaki Seisaku, Tie Jian, Haseba Takeshi, Cui Fanlai, Ohno Youkichi, Isobe Eiji, Isahai Isamu, Tsutsumi Hirofumi
Division of Legal Medicine, Department of Social Medicine, Nihon University School of Medicine, 30-1 Oyaguchi Kamicho, Itabashiku, Tokyo 173-8610, Japan.
Division of Legal Medicine, Department of Social Medicine, Nihon University School of Medicine, 30-1 Oyaguchi Kamicho, Itabashiku, Tokyo 173-8610, Japan.
Leg Med (Tokyo). 2016 May;20:33-6. doi: 10.1016/j.legalmed.2016.03.005. Epub 2016 Mar 11.
The detection of plankton DNA is one of the important methods for the diagnosis of drowning from postmortem tissues. This study investigated the quantities of picoplankton (Cyanobacteria) DNA in the lung, liver, kidney tissues and blood of drowned and non-drowned rabbits, and the sensitivity of detection of picoplankton DNA by polymerase chain reaction (PCR) detect for the diagnosis of death from drowning. For this purpose, the DNA of the 16S ribosomal RNA gene of picoplankton was quantitatively assayed from the tissues of drowned and non-drowned rabbits immersed in water after death. Each of the liver, kidney and lung tissues and blood were obtained from drowned and non-drowned rabbits. Picoplankton DNA in the tissues was extracted using the DNeasy® Blood & Tissue kit to determine the yield of picoplankton DNA from each tissue. TaqMan real-time PCR was performed for quantitative analysis of picoplankton DNA. Target DNA was detected in the liver, kidney and lung samples obtained from the drowned rabbits, while no picoplankton DNA was detected in the non-drowned rabbit tissues (except in lung samples). The results verified that direct PCR for the detection of picoplankton DNA is useful for the diagnosis of drowning. Although we observed seasonal changes in the quantity of picoplankton in river water, we were able to detect DNA from various organs of drowned bodies during the season when picoplankton were not the most abundant.
浮游生物DNA的检测是死后组织溺水诊断的重要方法之一。本研究调查了溺水和未溺水家兔肺、肝、肾组织及血液中微微型浮游生物（蓝细菌）DNA的含量，以及聚合酶链反应（PCR）检测微微型浮游生物DNA对溺水死亡诊断的敏感性。为此，对死后浸泡在水中的溺水和未溺水家兔组织中的微微型浮游生物16S核糖体RNA基因的DNA进行了定量分析。从溺水和未溺水家兔获取肝、肾、肺组织及血液。使用DNeasy®血液和组织试剂盒提取组织中的微微型浮游生物DNA，以确定每个组织中微微型浮游生物DNA的产量。采用TaqMan实时PCR对微微型浮游生物DNA进行定量分析。在溺水家兔的肝、肾和肺样本中检测到目标DNA，而在未溺水家兔组织（肺样本除外）中未检测到微微型浮游生物DNA。结果证实，直接PCR检测微微型浮游生物DNA对溺水诊断有用。尽管我们观察到河水中微微型浮游生物数量存在季节性变化，但在微微型浮游生物数量并非最多的季节，我们仍能够从溺亡尸体的各个器官中检测到DNA。
