微流控芯片对血管紧张素原基因多态性的研究
首发时间:2004-04-05
摘要:目的:以微流控芯片为平台,以激光诱导荧光为检测手段,旨在探索AGT基因-6G→A位点基因多态性在大连地区人群中的分布及与原发性高血压的关联,并为大规模人群基因多态性分析提供一高效,快速和灵敏的检测技术。方法:采用聚合酶链反应-限制性片段长度多态性(polymerase chain reaction restriction fragment length polymorphism,PCR-RELP)的方法,分别运用微流控芯片电泳与琼脂糖凝胶电泳对大连地区汉族人群103例正常人与123例高血压患者AGT基因(-6)位点的多态性进行分析。结果:1. 微流控芯片能在250s内对AGT基因PCR产物酶切片段进行芯片电泳分析,,酶切片段迁移时间的相对标准偏差(relative standard deviation ,RSD)为0.12%~0.26%<1%(n=6)。2.大连地区正常对照组与高血压组中AGT基因(-6)位点A等位基因均有较高的发生频率(0.7038,0.7073),突变位点多态性无统计学差异(χ2=0.024, P>0.05),另外与西藏人群相比,两地A、G等位基因频率分布差异亦无显著性(χ2=1.663,P>0.05)。结论:1. 微流控芯片同琼脂糖凝胶电泳相比,分析速度比凝胶电泳快,耗样量少,检测灵敏度高。2.大连地区汉族人群中AGT基因核心启动子区域-6位点等位基因A有较高的分布频率,但G/A的基因变异EH发病不相关联,此位点可能仅仅是一个基因多态性标志。
关键词: 微流控芯片、基因诊断、原发性高血压、血管紧张素原基因、多态性
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The study of angiotensinogen gene polymorphism by microchip electrophoresis
Abstract:bjective To investigate the AGT gene polymorphism distribution and the relationship between the AGT gene polymorphism and essential hypertension (EH) in Dalian Hans population using the new microchip-based method ,then,to develop a microchip-bases electrophoretic method for rapid detection of genotype. Method one hundred and twenty-three patients with EH and one hundren and three controls recruited from Dalian Hans population respectively were examined. The gene polymorphisms of AGT(G(-6)A) was determined by polymerase chain reaction –restriction fragment length polymorphism(PCR-RFLP) with microchip electrophoresis and agarose gel electrophoresis. Result (1) Microchip electrophoresis can separate PCR products of AGT gene within 250s and range of relative standard deviation (RSD) of the migration time was 0.12%-0.26%<1%(n=6).(2) The A allele frequency was very high in both groups(control:0.7038,hypertensive:0.7073), it was almost identical in the different groups(χ2=0.024, P>0.05), th
Keywords: Microchip electrophoresis ,Gene diagnosis ,Angiotensinogen , Essential Hypertension, Polymorphism
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