目的 建立一种适合临床常规检测胰岛素受体底物1 (IRS1)rs1801278基因多态性的实验方法.方法 随机收集2015年3月～2016年6月解放军第三○二医院200名健康体检者的外周血标本,设计针对IRS1 rs1801278基因多态性位点的特异性引物和荧光探针,通过TaqMan-MGB实时荧光聚合酶链反应法检测rs1801278基因多态性,以基因测序法(金标准)进行验证,同时初步探讨IRS1 rs1801278不同基因型在本研究人群中的分布频率.结果 TaqMan-MGB双荧光探针法鉴别IRS1 rs1801278的3种基因型,与基因测序法结果一致率为100％.IRS1rs1801278基因多态性中T/T、C/T和C/C基因型在本研究人群中的分布频率分别为0％、1％和99％.结论 根据单核苷酸多态性基因型特异性探针和引物所建立的TaqMan-MGB荧光探针法能快速有效地进行IRS1rs1801278基因多态性分型,结果可靠,操作易行,然而对中国汉族2型糖尿病患者个体化治疗效果的预测价值有限,不建议临床常规开展.%Objective To establish an experimental method suitable for clinical routine detection of the gene polymorphisms of insulin receptor substrate 1 (IRS1) rs1801278.Methods The peripheral blood samples of 200 healthy controls in 302 Military Hospital of China from March 2015 to June 2016 were collected,the specific primers and fluorescent probe aimed at the polymorphic sites of gene polymorphisms of IRS1 rs1801278 were designed,and the gene polymorphisms of rs1801278 were detected by the TaqMan-MGB real-time fluorescence polymerase chain reaction and validated by gene sequencing method (golden standard).The distribution frequencies of different genotypes of IRS1 rs1801278 in this population were discussed as well.Results The TaqMan-MGB two-color fluorescent probe method could identify 3 genotypes of IRS1 rs1801278,achieving 100％ consistence with gene sequencing method.The distribution frequencies of T/T,C/T and C/C genotypes of gene polymorphisms of IRS1 rs1801278 in this population were 0％,1％ and 99％,respectively.Conclusion The established TaqMan-MGB fluorescent probe method based on the specific probe and primer of single nucleotide polymorphisms is rapid and effective in identifying the genotypes of gene polymorphisms of IRS1 rs1801278,the results are reliable,and the operation is easy,but the predictive value for the individualized therapeutic effect of Chinese Han type 2 diabetes patients is limited,which is not recommended for clinical routine practice.
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