基于通用报告引物的5个Y-SNP复合扩增及其单倍型

Development of SNP Multiplexes Utilizing Universal Reporter Primers for Forensic Purposes

目的建立一种简便、经济、高效的SNP复合扩增体系,为法医学应用打下方法学基础。方法选择5个Y-SNP位点——IMS-JST164520、IMS-JST021354、IMS-JST003305、M119和M134,针对每一位点设计5′端带有通用报告引物(universal reporter primer,URP)的等位基因特异性引物,先利用等位基因特异性PCR技术扩增不同位点的等位基因片段,再利用荧光标记的URP扩增检测所有位点的等位基因。结果成功构建了5个Y-SNP荧光复合扩增体系,分型结果显示:同一SNP位点的两个不同等位基因表现为不同颜色的产物峰,不同SNP位点间等位基因片段长度不同。5个Y-SNP在武汉汉族群体中的单倍型多样性为0.8655。结论基于URP的SNP复合扩增体系具有简便、经济、高效的特点,具有较高的法医学应用价值。

Objective To establish a simple, fast and economical technique for multiplex-typing SNPs and to explore its potential forensic application. Methods Five Y-SNP loci（IMS-JST164520, IMS-JST021354, IMS-JST003305, M 119 and M134） were selected and the allele specific primers of each locus were designed with the universal reporter primers tailed at their 5＇ end. Alleles of these loci were amplified first by allele specific primers, then amplified by universal reporter primers tagged by fluorescent dye. Results A fluorescent-multiplex PCR system of the five Y-SNP loci was established. The typing results showed that two different colors of product peaks denoted two different alleles of a SNP locus, and the fragment sizes of alleles among different SNP loci were different. The haplotype diversity of these five loci was estimated to be 0.8655 in Wuhan Han population. Conclusion The multiplex-typing SNPs based on the universal reporter primers is a simple, fast, and economical technique, and may have good application value in forensic medicine.