【摘要】 目的 评价聚合酶链反应（PCR）结合熔解曲线（简称溶解曲线法）在高血压药物相关基因突变检测中的应用价值。方法 采用PCR熔解曲线法检测100例高血压患者5个高血压药物相关基因的单核苷酸多态性（SNP）位点[细胞色素P450同工酶2D6(CYP2D6)rs1065852、细胞色素P450同工酶2C9(CYP2C9)rs1057910、β1肾上腺素受体（ADRB1）rs1801253、血管紧张素Ⅱ受体1(AGTR1)rs5186、血管紧张素转换酶（ACE）rs1799752]，并与DNA测序结果进行比对。对PCR熔解曲线法的准确性、重复性、抗干扰能力进行评价。结果 PCR熔解曲线法与测序法对100例高血压患者的检测结果均一致，符合率为100%；检测5个基因位点Tm值的极值（R）均<0.5℃。血红蛋白（Hb）≤2 g/L、胆红素（Bil）≤342μmol/L、三酰甘油（TG）≤37 mmol/L对PCR熔解曲线法均无影响，Tm值的变异系数（CV）均≤5%。100例高血压患者中，AGTR1(c.1166A>C)及CYP2C9*3(c.1075A>C)纯合突变型各1例，CYP2D6*10基因型频率为39.5%,CYP2C9*3基因型频率为5.5%,ADRB1基因突变型C等位基因频率为62.0%,AGTR1基因突变型C等位基因频率为5.0%,ACE缺失型D等位基的频率为35.0%。结论 PCR熔解曲线法操作简便，对设备要求低，可作为检测降压药物相关基因突变的常规方法。更多还原

【Abstract】 Objective To assess the clinical application of polymerase chain reaction(PCR) melting curve method in determining hypertensive drug-related gene mutations. Methods Totally,100 hypertension patients were enrolled,and the single nucleotide polymorphism(SNP) sites of hypertensive drug-related gene mutations,including cytochrome P450 2D6(CYP2D6)rs1065852,cytochrome P450 2C9(CYP2C9)rs1057910,betaladrenergic receptor-1(ADRB1)rs1801253,angiotensin Ⅱ receptor 1(AGTR1)rs5186 and angiotensin-converting enzyme(ACE)rs1799752,were determined. DNA sequencing was performed. The accuracy,repeatability,antiinterference and the genotype frequency of PCR melting curve method were evaluated. Results The consistency of PCR melting curve method and DNA sequencing was 100%. The extremum values(R) of the Tm about 5 gene sites were all <0.5 ℃. Hemoglobin(Hb)≤2 g/L,bilirubin(Bil)≤342 μmol/L,triglyceride(TG)≤37 mmol/L had no effect on PCR melting curve method. The coefficient of variation(CV)of Tm was ≤5%. In 100 hypertension patients,there was 1 case of AGTR1(c.1166A>C) and 1 case of CYP2C9*3(c.1075A>C)homozygous mutations. The genotype frequencies of CYP2D6*10 and CYP2C9*3 were 39.5% and 5.5%,respectively. The genotype frequencies of C allele of mutant ADRB1,C allele of mutant AGTR1 and D allele of ACE deletion were 62.0%,5.0% and 35.0%,respectively. Conclusions PCR melting curve method is simple and has low requirements for equipments,and it can be used as a routine method for determining anti-hypertensive drug-related gene mutations.更多还原