【摘要】 目的探讨应用双重PCR-SSCP技术检测Wilson病(WD)患者基因突变的可行性,以期建立一种快速、高效的WD基因诊断方法。方法应用双重PCR技术扩增140例无亲缘关系的WD确诊患者和30例正常对照组的ATP7B基因第8、12外显子并行SSCP筛查,对出现异常泳动带型的外显子进行DNA测序确定突变位点和形式。结果正常对照组呈一种泳动带型,66例患者呈7种异常泳动带型,测序结果证实异常泳动带型均发生了突变,突变率占47.14%(66/140),其中Arg778Leu/G ln突变率为37.14%(52/140),Thr935M et突变率为12.86%(18/140)。结论应用双重PCR-SSCP技术检测WD基因第8、12等外显子是行之有效的方法,且可明显提高检测效率。更多还原

【Abstract】 Objective To establish a fast and effective gene diagnosis method for Wilson’s disease(WD) patients by double PCR-SSCP technology.Methods We amplificated exon8 and exon12 of ATP7B gene by double PCR from genomic DNA of 140 unrelated WD patients and 30 normal controls,then used SSCP technology to screen them.At last we identificated these patients’ mutation features by direct sequencing.Results No abnormality shift was found in 30 controls.In 140 patients,we found 7 types of abnormal mobility shifts in 66 cases(47.14%).In subsequent direct sequencing,mutation rate of Arg778Leu/Gln was 37.14%(52/140),and Thr935Met was 12.86%(18/140).Conclusion Double PCR-SSCP diagnosis technology is a effective method which can improve diagnosis rate for Wilson disease.更多还原