四引物法快速检测白细胞介素1A基因3'UTR插入/缺失多态性

Four primers method for the fast detection of insertion/deletion polymorphism in IL-1A gene 3'UTR

目的建立四引物扩增受阻突变体系聚合酶链反应(tetra-primer amplification refractory mutation system polymerase chain reaction,T-ARMS-PCR)检测白细胞介素1A基因3'UTR插入/缺失多态性的方法。方法使用试剂盒提取100例志愿者血样的基因组DNA,设计四引物检测白细胞介素1A基因3'UTR插入/缺失多态性的引物,对PCR的反应体系和扩增条件优化后对100例样本分型。DNA测序验证四引物法的准确性。结果 100例样本中Del/Del基因型39例,Ins/Ins基因型10例,Ins/Del基因型51例。DNA测序与四引物法的分型结果完全一致。结论四引物法可以实现白细胞介素1A基因3'UTR插入/缺失多态性的快速、简便、低成本检测。

Objective To establish a method for detection of insertion/deletion polymorphism in IL-1A gene 3'UTR by tetra-primer amplification refractory mutation system polymerase chain reaction(T-ARMS-PCR).Methods Genomic DNA was extracted from blood samples of 100 volunteers using DNA extraction kits.Four primers were designed to detect the insertion/deletion polymorphism in IL-1A gene 3'UTR.After the optimization of PCR conditions,100 samples were genotyped.The results of T-ARMS-PCR were verified by DNA sequencing.Results Among 100 samples,39 samples were Del/Del genotype,10 samples were Ins/Ins genotype,and 51 samples were Ins/Del genotype.The results of DNA sequencing were consistent with the results of T-ARMS-PCR.Conclusion The T-ARMS-PCR method is a simple,rapid and low-cost genotyping method for the detection of insertion/deletion polymorphism in IL-1A gene 3'UTR.