A diagnosis of the death by drowning using a molecular biological approach

目前，肺和其他器官以及骨髓中的硅藻鉴定对于溺水的诊断仍然是确凿的。然而，该方法存在一些问题，如硅藻的破坏和/或丢失，以及无机晶体的出现，干扰了显微镜观察。为了解决这些问题，本研究试图建立一种新的简便的方法--16S核糖体DNA(RDNA)的聚合酶链式反应分析，用于海洋细菌溺水的诊断。我们设计了海洋细菌(Listonella Angillarum)16S rDNA的特异性引物区。用所设计的引物扩增出100fg的鳗李斯特氏菌模板DNA。未检出各种细菌(大肠埃希菌、粪肠球菌、肺炎克雷伯菌、粘质沙雷氏菌、金黄色葡萄球菌、铜绿假单胞菌、枯草杆菌、普通变形杆菌、化脓性链球菌、流感嗜血杆菌)的PCR产物。当海水与人血混合时，从血样中检测到特异的聚合酶链式反应产物。这些结果表明，海洋细菌的分子生物学检测对溺水的诊断是有用的。

The diatom identification in lungs and in other organs, as well as in the bone marrow, remains at the present time quite conclusive for the diagnosis of drowning. However, the method has several problems such as destruction and/or loss of diatoms, and an appearance of inorganic crystals which disturbs microscopic observation. To solve these problem, in this study we attempted to establish a new easy method, PCR analysis of the 16S ribosomal DNA (rDNA), to diagnose drowning using marine bacteria. We designed the specific primers regions in 16S rDNA of the marine bacteria (Listonella anguillarum). The 100 fg of template DNA from Listonella anguillarum was amplified by the primers. PCR products from the various bacteria (Escherichia coli, Enterococcus faecalis, Klebsiella pneumoniae, Serratia marcesceus, Staphylococcus aureus, Pseudomonas aeruginosa, Bacillus subtilis, Proteus vulgaris, Streptococcus pyogens, Hemophilus influenza) were not detected. When the sea-water was mixed with human blood, the specific PCR products were detected from the blood sample. These results suggested that molecular biological detection of the marine bacteria is useful to diagnose drowning.