变性高压液相色谱分析在中国家族性CADASIL患者基因筛查中的应用

目的:探讨变性高压液相色谱分析(DHPLC)在伴皮质下梗死及白质脑病的常染色体显性遗传性脑动脉病(CADASIL)患者诊断中的方法和价值。方法:运用DHPLC及DNA直接测序技术,对CADASIL家系先证者、家系成员14例(CADASIL组)及健康对照者100例(对照组)进行Notch3基因检测。结果:CADASIL组发现3种致病性突变,其中Cys134Tyr为新的突变类型,同时发现15种多态类型;对照组中未发现突变。结论:DHPLC技术在筛查Notch3中起重要作用,但在筛查时应进行多个柱温的测试,温度调换以2℃为宜。

Application of Denaturing High Performance Liquid Chromatography in Genetic Diagnosis in Chinese Patients with Cerebral Autosomal Dominant Arteriopathy with Subcortical Infarcts and Leukoencephalopathy

Objective: To observe the significance of denaturing high performance liquid chromatography(DHPLC) in genetic diagnosis in chinese patients with cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy(CADASIL).Methods: DHPLC and DNA sequencing were used to identify the pathogenetic mutations in Notch3 gene in 4 probands and 10 family members from 3 unrelated Chinese CADASIL families and 100 healthy controls.Results: Three heterozygous missense mutations in exons 3 and 4 were found among the families but not in 200 chromosomes of controls.The Cys134Tyr was a novel mutation that has not been reported previously.There were also 15 polymorphisms found.Conclusion: DHPLC was a sensitive method to screen for mutations in Notch3 gene,but attention should be paid to optimize the oven temperature with the fluctuation of two degrees.