四引物扩增受阻突变体系快速检测Wilson病基因Arg778Leu突变  

Rapid identification of Arg778Leu gene mutation in Wilson's disease by 4-primer ARMS-PCR

作　　者：王平[1] 吴健民[1] 崔天盆[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院检验科,武汉430022

出　　处：《临床检验杂志》2007年第2期123-125,共3页Chinese Journal of Clinical Laboratory Science

基　　金：湖北省科技厅攻关课题(No:2004AA301C16)

摘　　要：目的建立一种不需要限制性片段长度多态性或直接测序的新方法检测肝豆状核变性(WD)病人突变热点ATP7B基因的Arg778Leu突变基因型。方法用四引物扩增受阻突变体系聚合酶链反应(tetra-primer amplification refractory mutation system-PCR,tetra-primer ARMS-PCR)检测47例WD患者和30例正常对照的ATP7B基因Arg778Leu突变,并用测序进行验证。结果47例WD患者中检出Arg778Leu纯合突变4例,杂合突变14例,总检出率38.3%(18/47);30例正常对照未发现突变;DNA测序结果与四引物ARMS-PCR结果完全一致。结论ATP7B基因Arg778Leu突变是中国人WD突变热点;四引物ARMS-PCR法检测Arg778Leu突变有快速、简便、准确的优点,可以区分等位基因是否纯合,适于大样本的人群筛查。此法也可用于检测其他点突变。Objective The aim of this study was to establish a new method for genotyping ATP7B Arg778Leu gene mutation that does not require RFLP PCR or sequencing. Method 4-primer amplification refractory mutation system （ARMS）-PCR was performed to screen the Arg778Leu mutation in 47 unrelated Wilson＇ s disease （WD） patients and 30 unrelated healthy controls. Direct sequencing was used to confirm the specific amplification products. Results PCR products were visualized on agarose gel electrophoresis. Among the 47 W D patients,4 were homozygous and 14 were heterozygous for this mutation. The total mutation rate was 38.3% （18/47）. The results of direct sequencing completely consisted with the results of 4-primer ARMS-PCR. Conclusions The ATP7B Arg778Leu gene mutation is a hot spot for the research of Chinese WD patients. 4-primer ARMS-PCR is a fast convenient and accurate method for typing mutation in high throughput population screening. This approach can be used to detect other point mutations.

关 键 词：肝豆状核变性 四引物扩增受阻突变体系 基因诊断 

分 类 号：Q78[生物学—分子生物学]