检测藻类16SrDNA特异性片段在溺死诊断中的应用

发布时间：2018-05-27 02:20

本文选题：法医病理学 + 溺死　；参考：《南方医科大学学报》2015年08期

【摘要】：目的建立藻类16Sr DNA特异性片段扩增方法,探讨其在溺死诊断中的应用价值。方法 35只实验兔随机分组:生前入水组(溺死组)15只,死后入水组(空气栓塞致死后入水)15只,对照组(空气栓塞死后不作处理)5只;微波消解-真空抽滤-电镜扫描法检测过的20例水中尸体肝脏样本20份(硅藻阳性14份,阴性6份)。7种已知藻(直链藻、菱形藻、针杆藻、舟形藻,铜绿微囊藻,小环藻,小球藻)作为对照。提取组织样本及藻类DNA,扩增产物银染显带。结果生前入水组肺、肝、肾检出率分别为100%,86%,86%;死后入水组肺、肝、肾检出率分别为13%,0%,0%。对照组肺、肝、肾藻类未检出。生前入水组与死后入水组各种脏器中藻类检出率差异显著(P0.05)。20份经微波消解-真空抽滤-电镜扫描法检测的水中尸体肝脏样本中,使用本方法15份样本结果为阳性(包括1份硅藻阴性样本)。7种藻类DNA扩增结果为阳性。结论本文所建立的PCR法检测藻类16Sr DNA灵敏度高,可对多种溺死藻类同时检测,有较好的应用前景。
[Abstract]:Objective to establish a 16Sr DNA specific fragment amplification method for algae and to explore its application value in the diagnosis of drowning. Methods Thirty-five rabbits were randomly divided into three groups: drowning group (15 rats), postmortem group (15 rabbits died of air embolism, 15 rabbits died of air embolism) and control group (5 rabbits died of air embolism). A total of 20 liver samples (14 positive from diatoms and 6 negative from 6 species of known algae) were detected by microwave digestion, vacuum filtration and scanning electron microscopy in 20 cases of dead bodies in the water (straight chain algae, diamondophyta, Needles, scaphoid, microcystis aeruginosa, microcystis micronuginosa, Cyclocystis microcystis, Cyclocystis microcystis). Chlorella vulgaris) as control. Tissue samples and algae DNA were extracted and amplified by silver staining band. Results the positive rates of lung, liver and kidney in water group were 100% and 100%, respectively, and the positive rates of lung, liver and kidney in postmortem water group were 130% and 0% respectively. In control group, no algae were detected in lung, liver and kidney. There was a significant difference in the detection rate of algae between antemortem group and postmortem group. There were 20 samples of liver detected by microwave digestion, vacuum filtration and electron microscope. The results of DNA amplification were positive in 15 samples (including 1 negative sample of diatoms). Conclusion the PCR method developed in this paper has high sensitivity for detection of algae 16Sr DNA and can be used for simultaneous detection of many kinds of drowning algae.
【作者单位】：南方医科大学法医学系;广州市刑事科学技术研究所//公安部法医病理学重点实验室;广东省法医遗传学重点实验室;
【基金】：公安部科技强警基础工作专项项目(2014GABJC020)
【分类号】：R649.3;D919

【参考文献】