16个Y-STR基因座快速复合扩增体系的构建研究

Study on Construction of a 16-Plex Rapid Y-STR Amplification System

为了构建一套16个Y-STR基因座的快速复合扩增体系,用于满足公安的实战时效性需求,文中选择DYS19、DYS385a/b、DYS390、DYS391、DYS392、DYS393、DYS438、DYS439、DYS437、DYS448、DYS456、DYS458、DYS635、Y_GATA H4和DYS447共16个基因座,将Roche FastStart Taq DNA Polymerase体系与ProPlex热循环仪结合,以法医DNA标准品9948为实验模板,从扩增条件、扩增体积、缓冲液选择、DNA聚合酶用量、基因座间的平衡性调整、快速扩增程序的优化等方面进行一系列复合扩增实验,比较不同条件下等位基因的丢失、峰高、基因座间峰的均衡性及非特异峰。结果发现,该快速体系30 min即可获得样本全部16个Y-STR基因座分型,且各等位基因间均衡性良好,无非特异性峰。以上信息初步表明建立的16个Y-STR基因座的快速复合扩增体系可明显提高样品检测效率,对实战中一些时效性案件具有一定的实际意义。

To establish a 16-plex rapid Y-STR amplification system for the actual needs of public security, sixteen STR loci, including DYS19, DYS385a/b, DYS390, DYS391, DYS392, DYS393, DYS438, DYS439, DYS437, DYS448, DYS456, DYS458, DYS635, Y_GATA H4 and DYS447, were chosen based on the cur- rent Y-STR genotyping for development of the rapid PCR. Roche FastStart Taq DNA Polymerase and DNA standard sample 9948 were used during an amplification and optimization process. The 16-plex rapid Y- STR amplification system was achieved by performing various experiments, including choosing amplification conditions and the volume of DNA polymerase, and adjusting interlocus balance, thermal cycling parameters and the volume of rapid PCR system, reaction buffers and a lot of additives. The results showed that, through this 16-plex rapid Y-STR amplification system, all 16 samples of Y-STR loci could be obtained in 30 min- utes, with a good balance between the alleles and without nonspecific amplification. It proved that the rapid recombination amplification system of 16 Y-STR sites can improve the efficiency of sample detection and is of practical significance for the timeliness of special cases in public security.