Wilson病基因突变的检测和分析

Identification and Analysis of Mutations in Patients with Wilson Disease

目的:研究Wilson病(WD)基因突变的类型和发生情况,探讨WD基因诊断的意义和方法。方法:采用变性高效液相色谱(DHPLC)及单链构象多态性(SSCP)方法结合DNA序列分析对12个无亲缘家系13例患者及22位亲属进行ATP7B基因外显子5、8、12突变的检测。结果:35例DNA标本确认6种基因突变,其中包括3种错义突变(R778L,D756N,R952K),1种缺失突变(2790del3),2种同义突变(2310C→G,2850G→C)。在13例患者中,发现6例R778L突变,2例D756N突变,R952K和2790del3各1例,均为杂合型突变,涉及10个等位基因,本组检出率为38.5%(10/26)。结论:PCR-SSCP,DHPLC是WD基因突变筛查较为敏感而特异的方法,且二者各具特点。经检索2790del3是ATP7B基因新突变类型。R778L是中国人最常见的突变类型。

Objective： To determine mutation characters and distributions of ATP7B gene and explore significance and method of gene diagnosis in Wilson disease （WD） patients. Methods： Thirty five individuals （ 13 WD patients） from 12 no kinship WD families were subjected to polymerase chain reactions （PCR） for exons 5,8 and 12 of ATP7B gene. PCR products were analyzed by SSCP,DHPLC and by direct sequencing. Results： Six mutations were identified by DNA sequencing, including three missense mutations （R778L, D765N and R952K）, one deletion （2790del3） and two polymorphisms （2310C→G,2850G→C）. The mutation R778L occurred in 6 patients and they were all heterozygous. Conclusion： The methods of PCR-SSCP and DHPLC are dependable tool for screening ATP7B gene mutations. The 2790del3 is a novel mutation of ATP7B found in WD patients. The mutation R778L is the most common mutation of ATP7B gene in Chinese WD patients.