用荧光PCR对中国人肝豆状核变性进行早期诊断及携带者检测

Using fluorescence PCR analysis for early diagnosis and carriers detection of Chinese Wilson's disease

目的 应用荧光聚合酶链反应技术对中国人肝豆状核变性 (Wilson's disease,WD)进行早期诊断和携带者检测。方法 在 6 6例 WD患者、5 5名健康的家系成员和 30例其他疾病患者 (作为非 WD对照组 )中 ,检测了中国人 WD基因的突变热点 Arg778L eu;随机抽取 3例样本 (2例来自 WD患者组 ,1例来自对照组 )进行 DNA测序以验证结果。结果 在 6 6例 WD患者中检出 5例 Arg778L eu突变纯合子和 2 1例突变杂合子 ,总检出率为 39.4% ;在 5 5名 WD家系成员中检出了 12例杂合子 ,并确定其中 11例是 WD致病基因携带者而不是症状前患者 ;DNA测序结果与荧光 PCR结果一致。结论 8号外显子 Arg778L eu突变是中国人 WD的高频突变点 ,荧光 PCR具有简便快速、准确、检出率高等优点目的 应用荧光聚合酶链反应技术对中国人肝豆状核变性 (Wilson's disease,WD)进行早期诊断和携带者检测。方法 在 6 6例 WD患者、5 5名健康的家系成员和 30例其他疾病患者 (作为非 WD对照组 )中 ,检测了中国人 WD基因的突变热点 Arg778L eu;随机抽取 3例样本 (2例来自 WD患者组 ,1例来自对照组 )进行 DNA测序以验证结果。结果 在 6 6例 WD患者中检出 5例 Arg778L eu突变纯合子和 2 1例突变杂合子 ,总检出率为 39.4% ;在 5 5名 WD家系成员中检出?

Objective To diagnose the pre symptomatic cases of Chinese Wilson's disease(WD) and detect the potential carriers by using fluorescence PCR technique. Methods Screening the high frequency spot Arg778Leu of WD gene mutation in 66 Chinese WD patients, 55 healthy family members and 30 controls, and selecting 3 random samples (2 from WD patients, 1 from controls) for DNA sequencing to testify the accuracy of fluorescence PCR. Results Among 66 Chinese WD cases, 5 were found homozygous for mutation of Arg778Leu and 21 were compound heterozygous; the gene mutation rate was totally 39.4%.Out of 55 healthy family members, 12 were found heterozygous, and it was confirmed that 11 of these 12 individuals were WD gene carriers but not pre symptomatic patients. The results of direct DNA sequencing consisted with those results detected by fluorescence PCR. Conclusion The Arg778Leu mutation in exon 8 is the high frequency spot of Chinese Wilson's disease gene. Fluorescence PCR analysis is a rapid, accurate gene diagnostic method and demonstrates a high detecting rate.