四个STR基因座荧光标记复合扩增的研究及其法医学应用

发布时间：2018-02-01 18:05

本文关键词： 法医学 DNA分型短串联重复序列(STR) 聚合酶链式反应(PCR) 复合扩增荧光标记毛细管电泳荧光检测自动分析法群体遗传学　出处：《四川大学》2007年硕士论文　论文类型：学位论文

【摘要】： 目的本课题旨在建立已有群体遗传学数据的STR基因座D11S4951、D7S3062、GATA168F06和D10S1426基因座的荧光标记复合扩增体系。按照美国DNA分析方法技术工作组(The Technology Working Group on DNA Analysis Methods，TWGDAM)的指导方案进行法医学实用性研究。方法要首先对多个本实验室做过群体遗传学研究的STR基因座进行筛选，从中获得四个多态性好、个人识别机率高，扩增产物长度片段有区别的四个STR基因座，即D11S4951、D7S3062、GATA168F06、D10S1426，利用传统的复合扩增方法，ABI-310毛细管电泳仪进行荧光标记复合扩增的研究。并对复合扩增体系的灵敏度和准确度，不同扩增循环次数和热启动下复合扩增的效果等进行了研究。按照TWGDAM指导方案的基本要求，对该体系进行了法医学实用性研究，包括基因座的种属特异性，组织同一性及可重复性。结果成功建立了D11S4951、D7S3062、GATA168F06和D10S1426的荧光标记复合扩增体系。而且，这一扩增体系条件易于优化，采用国产Taq酶，不需要热启动进行扩增，即可得到满意的扩增产物和分型结果。相对于国外昂贵的试剂盒，是一种成本低廉但效率高的STR基因座复合扩增体系。法医学应用性研究表明，，该体系的检测灵敏度为0.25ng模板DNA，四个STR基因座具有较高的种属特异性，对同一个体的不同组织检测结果一致，对常见基质上的检材及混合样本具有检测能力。对15个实际检案样本的检测证明，该体系能用于法医学个人识别和亲权鉴定。结论D11S4951、D7S3062、GATA168F06和D10S1426的荧光标记复合扩增体系可应用ABI-310检测平台，获得可靠的分型结果。法医学应用性研究证明，该体系扩增条件易于优化、成本低廉、分型结果稳定，灵敏度和准确度高，种属特异性好，重复性好，对陈旧斑痕具有检测能力，能够对常见基质上的斑痕以及混合斑痕正确分型，与常见的动物及微生物无交叉反应，可以应用于法医学实际检案。
[Abstract]:Objective to establish a STR locus D11S4951 and D7S3062 with population genetic data. Fluorescent multiplex amplification system for GATA168F06 and D10S1426 loci. Technical working Group on DNA Analysis in the United States (. The Technology Working Group on DNA Analysis Methods. The guidance program of TWGDAM was used to study the practicability of forensic medicine. Methods STR loci which had been studied in population genetics in our laboratory were screened and four polymorphisms were obtained. There were four STR loci with different length fragments of the amplified products, D11S4951, D7S3062, GATA168F06 and D10S1426. The conventional multiplex amplification method (ABI-310 capillary electrophoresis instrument) was used to study the fluorescence labeling multiplex amplification. The sensitivity and accuracy of the multiplex amplification system were also studied. The effects of multiple amplification under different amplification cycles and hot start were studied. According to the basic requirements of TWGDAM guidance scheme, the practicability of forensic medicine was studied. The results showed that D11S4951D7S3062 was successfully established. GATA168F06 and D10S1426 multiplex amplification system, and this amplification system conditions are easy to optimize, using domestic Taq enzyme, without the need for hot start to amplify. Satisfactory amplification products and typing results can be obtained. Compared with expensive foreign kits, it is a low cost but high efficiency STR locus complex amplification system. Forensic application studies show that. The sensitivity of the system is 0.25 ng template DNA. The four STR loci have higher species-specificity, and the results are consistent for different tissues of the same individual. This system can be used in personal identification and paternity identification of forensic medicine. Conclusion D11S4951. The fluorescent multiplex amplification system of D7S3062Gata 168F06 and D10S1426 can be detected by ABI-310. The results of forensic application showed that the system was easy to optimize, low cost, stable typing results, high sensitivity and accuracy, good species specificity, good reproducibility. It has the ability to detect the old marks, can correctly type the stains and mixed stains on common substrates, and has no cross reaction with common animals and microorganisms, so it can be used in forensic medical practice.
【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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