A study on detection of rbcL gene for diatoms involved in drowning
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A study on detection of rbcL gene for diatoms involved in drowning
Journal of Chongqing Medical UniversityVol. 43, Issue 1, Pages: 98(2018)
作者机构:
1. 广东工业大学轻工化工学院生物工程系,广州,510006
2. 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,广州,510000
3. 南方医科大学法医学院,广州,510515
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Published:2018,
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Peng Fan, Liu Chao, Xu Quyi, et al. A study on detection of rbcL gene for diatoms involved in drowning. [J]. Journal of Chongqing Medical University 43(1):98(2018)
DOI:
Peng Fan, Liu Chao, Xu Quyi, et al. A study on detection of rbcL gene for diatoms involved in drowning. [J]. Journal of Chongqing Medical University 43(1):98(2018)DOI:
A study on detection of rbcL gene for diatoms involved in drowning
Objective:To establish the PCR method with capillary electrophoresis(PCR-CE) for the detection of rbcL gene of diatoms involved in drowning based on the specific primers for diatoms rbcL gene. Methods:With 29 microbes probably involved with drown-ing,a PCR-CE detection system with the primer for diatom rbcL gene labelled with fluorescent was established to verify the specificity and stability. The effectiveness of the PCR-CE method for the diatoms rbcL gene detection in corpses samples was evaluated by com-paring the results of MD-VF-Auto SEM. Results:Positive results(size=197 bp) were achieved with 6 diatoms species(Navicula sp.,Nitzschia sp.,Cyclotella sp.,Melosira varians,Synedra radians,Skeletonema) by the PCR-CE method,by contrast with negative out-comes for other species. The sensitivity of the above 6 species of diatoms DNA were 0.502,0.117,0.029,0.042,0.275,0.215 ng,respectively(20 μL PCR system). 35 cases of corpses(3 corpses land deaths,2 waterborne corpse cases of non-drowning,30 drown-ing cases) were detected,the detection rate of diatoms in the lungs,liver and kidney of the corpses was 100%,63.3% and 53.3%,respectively. The total positive rate was 83.3%. When the diatom content >10/10 g,the total positive rate was 100% by MD-VF-Auto SEM method. The total positive rate was and 92.6% by PCR-CE,there was no statistical difference( ?字2=2.077,P=0.491),otherwise,there was statistical difference(P<0.05). Conclusion:Because of less sample consumption,the PCR-CE method based on the ND-rbcL primer has a good prospect in forensic application for diatom detection,and leads to easy popularization in drowning diagnosis.