CODIS系统三个miniSTR荧光标记复合扩增的构建及法医学应用

发布时间：2018-05-29 11:35

  本文选题：miniSTR + 短串联重复序列　； 参考：《四川大学》2007年硕士论文

【摘要】： 目的对THO1、TPOX、CSF1PO三个基因座PCR-miniSTR和普通引物PCR-STR的结果进行一致性检验；构建THO1、TPOX、CSF1PO三个基因座miniSTR荧光标记复合扩增体系；对该荧光标记复合扩增体系进行法医学应用性研究。方法参照Bulter等对THO1、TPOX、CSF1PO三个基因座miniSTR的引物设计，选用本教研室提供的123例无血缘关系个体的血样，进行THO1、TPOX、CSF1PO三个基因座PCR-miniSTR银染检测，检测结果与商品化试剂盒的检测结果进行一致性检验；采用国产DNA聚合酶构建miniSTR荧光标记复合扩增体系，用美国ABI-310基因分析仪进行检测；对该荧光标记复合扩增体系的灵敏度、准确性、种属特异性、不同退火温度下复合扩增的效果、陈旧血痕DNA的检测等进行研究。结果123例无血缘关系个体的THO1、TPOX、CSF1PO三个基因座PCR-miniSTR银染检测结果与商品化试剂盒检测结果一致；成功建立了THO1、TPOX、CSF1PO三个基因座miniSTR荧光标记复合扩增体系，该体系的的退火温度范围较宽，以60℃为最佳，检测灵敏度为0.25ng模板DNA，，扩增准确性高，具有较高的种属特异性，可以应用于陈旧血痕DNA检测。结论THO1、TPOX、CSF1PO三个基因座PCR-miniSTR结果可以与该三个基因座普通引物PCR-STR的结果进行比对，具有数据库兼容性；该荧光标记复合扩增体系可应用于ABI-310检测平台，扩增灵敏度高，条件易于优化、分型结果稳定，种属特异性好，可应用于法医学实际检案。
[Abstract]:Objective to verify the consistency between the PCR-miniSTR of TPOXG CSF1PO and the PCR-STR of common primers, to construct a fluorescent multiplex amplification system for miniSTR of TPOXG / CSF1PO, and to study the forensic application of the system. Methods according to the primer design of miniSTR of three loci of THO1TPOXG CSF1PO, the blood samples of 123 unrelated individuals from our teaching and research department were used to detect the PCR-miniSTR silver staining of the three loci of THO1, TPOX, CSF1PO. The detection results were consistent with those of commercial kits. The miniSTR fluorescent labeling multiplex amplification system was constructed by domestic DNA polymerase and detected by American ABI-310 gene analyzer. The sensitivity of the fluorescent labeling multiplex amplification system was analyzed. Accuracy, species specificity, multiple amplification at different annealing temperatures, and DNA detection of old blood marks were studied. Results the results of PCR-miniSTR silver staining of the three loci of THO1 and TPOXF1PO in 123 unrelated individuals were in agreement with the results of commercial kit, and the fluorescent multiplex amplification system was successfully established for the miniSTR labeling of the three loci of THO1, TPOX, CSF1PO, and the annealing temperature of the system was wide. At 60 鈩

本文编号：1950741