TaqMan探针技术用于X-SNP位点的分型

X-SNP Genotyping Using the TaqMan Probe Technology

目的建立一种基于TaqMan探针技术的快速、准确且经济的实时荧光PCR方法,用于检测X染色体上的单核苷酸多态性(single nucleotide polymorphism,SNP)。方法选择X染色体上的13个SNP位点(X-SNP),针对每个位点分别设计1对PCR引物和TaqMan探针,进行实时荧光PCR扩增,对X-SNP位点进行分型。结果13个位点均符合Hardy-Weinberg平衡;多态信息含量分布为0.3497～0.3750,杂合度为0.4537～0.5021。建立的方法能够用于X-SNP位点的基因分型,检测结果与DNA测序结果完全一致。结论基于TaqMan探针技术的等位基因特异的实时荧光PCR方法灵敏、简单、快速,可实现对X-SNP位点的分型检测;所选择的13个X-SNP位点具有高信息量,在法医遗传学中具有潜在的应用价值。

Objective To develop a rapid, accurate and economical real time fluorescence PCR method with TaqMan probe technology to detect the X chromosome single nucleotide polymorphism（X-SNP）. Methods Taq- Man probes and polymerase chain reaction primers were respectively designed according to the 13 X-SNP. Then, the X-SNP were genotyped after the amplification by real time fluorescence PCR. Results All the loci follow the Hardy-Weinberg equilibrium. The polymorphic information content for 13 distinct loci varied between 0.3497 and 0.375 0 while the heterozygosity ranged from 0.453 7 to 0.502 1. A real time fluorescent PCR method based on TaqMan probe was successfully developed and the resuhs were accordant with those analyzed by DNA sequencing of the 13 X-SNP. Conclusion The allele specific real time fluorescence PCR based on TaqMan probe is a sensitive, simple technology and suitable for rapid analysis of X- SNP. All the loci show highly polymorphic and may be potential in forensic genetics.