三个miniSTR遗传标记及其荧光标记复合扩增的法医学应用研究

发布时间：2018-04-10 21:23

本文选题：法医遗传学 + 微小片段短串联重复序列(miniSTR)　；参考：《四川大学》2007年硕士论文

【摘要】： 目的miniSTR技术是当今法医物证学研究高度降解检材的热点，本课题旨在研究3个miniSTR基因座(D10S1248，D14S1434和D22S1045)在中国成都汉族人群中的特征，构建复合扩增荧光标记检测系统及评估其法医学应用价值。方法使用Coble的引物，在中国成都200个无关汉族个体中进行3个miniSTR基因座PCR扩增，扩增产物使用聚丙烯酰胺凝胶电泳分离，银染法显色，以获得群体基本数据。建立这3个miniSTR基因座的复合扩增体系，利用荧光复合扩增技术扩增，ABI PRISM~(?) 310基因分析仪对产物进行检测，GeneScan3.7及Geno Typer3.7软件分析结果；并按照美国DNA分析方法技术工作组(Scientific Working Group on DNA Analysis Methods，，SWGDAM)的指导方案进行法医学应用性研究，用不同的组织、不同的介质、常见的动物对复合扩增系统进行测试，用实际案例进行验证。结果获得了3个miniSTR基因座的基因频率、个人识别率、非父排除率等群体遗传学数据，在D10S1248基因座观察到了新的等位基因8，在D14S1434基因座发现了不同的重复序列；通过采用复合扩增方法，建立了D10S1248、D14S1434和D22S1045荧光标记复合扩增体系。法医应用性研究证明该体系与12种常见动物和微生物无交叉反应；可对2ng的微量DNA模板分型；对同一个体的不同组织检测结果一致；能够对常见介质上的斑痕检材正确分型；对放置20年的陈旧血痕仍然可分型；应用于案例检测，结果与鉴定结论一致。结论3个miniSTR遗传标记(D10S1248，D14S1434和D22S1045)在中国人群中有较高的多态性及较高的个人识别率，在不同的人群中存在遗传差异。建立了这3个miniSTR基因座的荧光标记复合扩增体系miniplex，该体系可运用于荧光标记毛细管电泳激光自动分析检测平台，具备法医学实践应用的可行性，为分析DNA降解检材提供了一个有效的途径。
[Abstract]:Objective to study the characteristics of three miniSTR loci (D10S1248, D14S1434 and D22S1045) in Chengdu, China.To construct a multiplex amplification fluorescent marker detection system and evaluate its forensic application value.Methods the primers of Coble were used to amplify three miniSTR loci from 200 unrelated Han Chinese individuals in Chengdu, China. The amplified products were separated by polyacrylamide gel electrophoresis and colored by silver staining to obtain the basic population data.To establish the multiplex amplification system of the three miniSTR loci, and to detect the products by using the fluorescence multiplex amplification technique to detect the products with GeneScan3.7 and Geno Typer3.7 software.In accordance with the guidance of Scientific Working Group on DNA Analysis methods (SWGDAM), the United States DNA Analytical Methodology Technical working Group conducted forensic application research, and tested the multiplex amplification system with different tissues, different media and common animals.An actual case is used to verify the problem.Results the population genetic data of three miniSTR loci, such as gene frequency, personal identification rate and non-paternal exclusion rate, were obtained. New alleles 8 were observed in D10S1248 loci and different repeat sequences were found in D14S1434 loci.The multiplex amplification system of D10S1248, D14S1434 and D22S1045 was established by using the method of multiplex amplification.It can still be classified for 20 years old blood marks, and the results are consistent with the identification results.Conclusion three miniSTR markers D10S1248, D14S1434 and D22S1045) have higher polymorphism and higher personal identification rate in Chinese population, and there are genetic differences among different populations.The fluorescent multiplex amplification system of these three miniSTR loci was established. The system can be used in fluorescence labeling capillary electrophoresis laser automatic analysis and detection platform, which has the feasibility of forensic practice.It provides an effective way for the analysis of DNA degradation samples.
【学位授予单位】：四川大学
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：D919

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