建立一种实时荧光PCR对维生素D受体基因单核苷酸多态性快速分型

Establishment of a Real-Time Fluorescent PCR for Single Nucleotide Polymorphism of Vitamin D Receptor Gene

目的:建立实时荧光PCR快速检测维生素D受体(VDR)基因rs2228570、rs1544410、rs7975232位点多态性。方法:以VDR基因的rs2228570、rs1544410、rs7975232三个SNP位点的变异碱基分别设计并合成特异性引物。11例体检健康儿童血液标本作为检测样本,采用实时荧光PCR方法检测样本中VDR基因型,采用Sanger法对检测结果测序验证。结果:利用所设计的特异性引物对人全血基因组中VDR进行SNP位点特异性扩增,根据7500 FAST实时荧光PCR仪的分析结果得到基因型,与测序结果比对后发现,11个样本采用实时荧光PCR方法基因分型结果与测序结果均一致。结论:实时荧光PCR快速检测VDR基因rs2228570、rs1544410、rs7975232位点多态性的方法或可用于临床VDR基因SNP快速分型。

Objective:To establish a real-time fluorescent PCR for rapid detection of polymorphisms of vitamin D receptor(VDR)gene rs2228570,rs1544410,rs7975232.Method:Based on the variant bases of three SNP sites of rs2228570,rs1544410 and rs7975232 of VDR gene,specific primers were designed and synthesized respectively.11 clinical blood samples were collected as templates,real-time fluorescent PCR method was used to detect the VDR genotype in the samples,and the detection results were sequenced and verified by Sanger method.Results:The specific primers were used to perform SNP site-specific amplification of VDR in the human whole blood genome.The genotypes were obtained based on the analysis results of the 7500 FAST real-time fluorescent PCR instrument.After comparison with the sequencing results,11 samples were found.The genotyping results using real-time fluorescent PCR method were consistent with the sequencing results.Conclusion:The real-time fluorescent PCR method for rapid detection of VDR gene rs2228570,rs1544410,rs7975232 loci polymorphism maybe used for clinical VDR gene SNP rapid typing.