Y染色体STR复合扩增及法医学可行性研究

发布时间：2018-06-18 03:05

本文选题：Y-STR + 复合扩增　；参考：《四川大学》2003年博士论文

【摘要】： 目的 Y染色体STR(Y-STR)呈单倍型父系遗传特征，不仅在人类学、古生物学等方面意义重大，而且在法医学的个人识别及亲子鉴定中亦具有重要而独特的应用价值。本课题旨在通过研究复合扩增技术，构建四个Y-STR基因座的复合扩增银染检测体系和复合扩增荧光检测体系，并对其进行法医学可行性研究和群体遗传研究。以期为法医Y染色体分型提供新的技术手段。方法选定四个Y染色体STR基因座，应用加尾序列引物设计策略设计的引物，构建四个基因座的Y-STR复合扩增体系，建立银染检测和荧光检测方法，依据DNA分析技术工作组(TWGDAM)指南进行法医学可行性研究和群体遗传研究。结果构建了四个Y-STR基因座：A10(Y-GATA-A10)，DYS434，DYS438，DYS439复合扩增体系(Y-A489-plex)；建立了Y-STR复合扩增银染检测方法和荧光检测方法。法医学可行性研究结果显示，敏感度检验最低模板量Y-A489-plex银染体系为0.5ng，Y-A489-plex荧光体系为0.125ng；分型结果稳定，重复性好，对陈旧斑痕具有检测能力，能够对常见介质上的斑痕正确分型，具有很好组织同一性、种属特异性，可以耐受女性成分的干扰，用于混合斑分析独具优势;可以用于实际检案。在100个样本中检出37个单倍型，单倍型变异度为0.9628，标准误为0.00429。结论本研究在国内外第一次将加尾序列引物设计策略引入 Y一STR复合扩增体系。证明该引物设计策略在Y一STR复合扩增研究中是切实可行的。加尾引物复合扩增策略还有很大的潜力，为大规模Y一STR复合扩增的研究提供了一个行之有效的方法。初步构建了四个Y一STR基因座的Y一A489一plex银染体系和 Y一A4Sg一plex荧光体系:并依据DNA分析技术工作组(TWGDAM) 指南进行了应用性研究。证明Y一A489一plex，适合于法医DNA分析，有较高的应用价值，，为法医Y染色体分型提供了新的技术手段。 Y一A489一plex体系采用的是国产的Taq酶。经与美国生产的金牌酶比较，无论是扩增效率还是特异性都无明显差异，为试剂国产化进行了有效的探索。
[Abstract]:Objective Y chromosome STR- Y-STR has haplotype paternal genetic characteristics, which not only has great significance in anthropology and paleontology, but also has important and unique application value in forensic personal identification and paternity test. The purpose of this study was to construct a multiplex amplification silver staining detection system and a multiplex amplified fluorescence detection system for four Y-STR loci, and to study the feasibility of forensic medicine and population heredity of the four Y-STR loci. In order to provide a new technique for forensic Y chromosome typing. Methods four STR loci of Y chromosome were selected. The Y-STR multiplex amplification system of the four loci was constructed by using the primers designed by adding tail sequence primers, and the silver staining and fluorescence detection methods were established. Forensic feasibility studies and population genetics studies were conducted in accordance with the TWGDAM guidelines of the working Group on DNA Analysis techniques. Results four Y-STR loci were constructed, and the multiplex amplification system of DYS434 (DYS438) DYS439 was constructed, and the silver staining and fluorescence detection methods of Y-STR multiplex amplification were established. The results of the feasibility study of forensic medicine showed that the minimum template quantity of sensitivity test was 0.5ngA489-plex silver staining system, the fluorescence system of Y-A489-plex fluorescence system was 0.125ng.The typing results were stable, reproducible, and had the ability to detect the old stains. Able to type correctly the stains on common media, with good tissue identity, species specificity, It can tolerate the interference of female component, and can be used for mixed spot analysis. 37 haplotypes were detected in 100 samples, the variation of haplotypes was 0.9628, and the standard error was 0.00429. Conclusion for the first time, the design strategy of tail sequence primer was introduced into Y-STR compound amplification system at home and abroad. It is proved that the primer design strategy is feasible in the study of Y-STR compound amplification. The strategy of adding tail primers has great potential, which provides an effective method for the study of large scale Y STR multiplex amplification. Y-A489 plex silver staining system and Y A4Sg plex fluorescent system were constructed with four Y STR loci. The guide conducts an application study. It is proved that Y A 489 complex is suitable for forensic DNA analysis and has high application value. It provides a new technical hand segment for forensic Y chromosome typing. Y-A489- plex system Domestic Taq enzyme was used. Compared with the gold enzyme produced in the United States, there was no significant difference in amplification efficiency or specificity. It is an effective exploration for the localization of reagents.
【学位授予单位】：四川大学
【学位级别】：博士
【学位授予年份】：2003
【分类号】：D919

【引证文献】