荧光标记复合扩增毛细管电泳法在SNP分型中的应用

Development of autosomal SNP fluorescent-multiplex by the method of fragment length discrepant allele specific PCR

目的采用荧光标记复合扩增毛细管电泳法,对辽南地区汉族人群13个SNP进行等位基因频率调查,并评价其法医学应用价值。方法选择13个双等位基因SNP,应用荧光标记片段长度差异等位基因特异性复合扩增SNP分型方法,对辽南地区汉族人群进行群体调查。结果每个SNP纯合子为单一产物峰,杂合子则为长度不同的两个产物峰。不同位点扩增产物长度不同,根据产物长度和产物峰数量进行SNP分型,其结果与直接测序完全一致。同时获得辽南地区汉族人群13个SNP等位基因频率。结论采用荧光标记复合扩增毛细管电泳法进行SNP分型,方法简单实用,在法医学个人识别领域具有较高的应用。

Objective To develop autosomal SNP fluorescent-multiplex PCR system, and investigate the allele frequencies of these 13 SNP loci in Southern Liaoning HaM population. Methods After selected 13 autosomal SNP loci, SNP fluorescent-multiplex system was developed based on the principle of fragment length discrepant allele specific PCR, and the allele frequencies of these SNP loci in Southern Liaoning HaM population were investigated using the established system. Results For the same SNP locus, the homozygote showed a single product peak, and the heterozygote showed two product peaks with different length. For the different SNP loci, the lengths of their PCR products were different. Therefore, we could genotype the 13 SNP loci simultaneously according to the length of products and the amount of product peaks, and the results were in accordance with that of direct sequencing. In the meanwhile, the allelic frequencies of these 13 SNP loci in Southern Liaoning HaM population were obtained. Conclusion The SNP fluorescent-multiplex system based on the of a high application fragment length discrepant allele specific PCR strategy is simple and economical, and is value in forensic medicine.