基于微流控芯片的72重单核苷酸多态性族群推断系统的构建

建立了常染色体单核苷酸多态性（SNPs）复合检测芯片体系，用于未知个体的族群来源推断。基于前期筛选的74-SNPs组合，采用竞争性等位基因特异性聚合酶链式反应（PCR）的原理构建SNPs的扩增体系，在微流控芯片的每个反应孔内完成一个SNP的检测，通过高通量PCR微流控芯片实现了其中72个SNPs的同步检测。芯片的扩增由平板PCR仪完成，反应孔的荧光信号通过激光共聚焦扫描仪检测，最终通过提取的荧光值进行结果分析。使用该芯片检测获得52份样本的SNPs分型，分型结果的准确率为100%。以57个人群的3628个样本为参考人群数据库，进行20份样本的族群来源推断，推断结果与样本的实际来源一致。本研究建立的常染色体72个SNPs微流控芯片体系可以有效地进行SNP多态性分析检测，基于参考数据库，20份检测样本族群推断的准确性为100%。

Construction of a microfluidic chip-based 72 plex single nucleotide polymorphisms ancestry inference system

The aim was to develop an autosomal single nucleotide polymorphism (SNP) multiplex detection chip system for the inference of the ancestry of unknown individuals. On the basis of the 74-ancestry informative SNP (AISNP) panel screened by our group, a multiplex SNP amplification and detection system, based on a microfluidic chip of competitive allele-specific polymerase chain reaction (PCR), was constructed. The allele-specific primers of each SNP were positioned into a single well on the microfluidic chip and each chip included 112 wells. For detection of SNPs, the chip was placed on the plate thermocycler to perform PCR. The fluorescence signal of the reaction in each well was detected by a confocal laser scanning instrument to evaluate the SNP genotypes of each sample. Fifty-two samples were used for SNP typing. The accuracy of typing results was 100%. Taking 3628 samples of 57 populations as the reference population database, the ancestry of 20 samples was inferred. The results were consistent with the actual sources of the samples. The autosomal 72-SNP microfluidic chip system established can accurately detect SNPs. The ancestry of tested individuals can be accurately inferred from the reference database.