应用微卫星DNA单体型检测Wilson病症状前患者及杂合子  

DETECTION OF PRESYMPTOMATIC PATIENTS AND HETEROZYGOTES WITH WILSON’S DISEASE BY USING HAPLOTYPES OF MICROSATELLITES

作　　者：王丽娟[1,2,3] 梁秀龄[1,2,3] 刘焯霖 沈福民 孟炜[1,2,3] 徐评议 潘锡榜[1,2,3] 陈彪 

机构地区：[1]广东省人民医院神经内科 [2]上海医科大学流行病学教研室 [3]美国帕金森氏病研究所

出　　处：《中华医学遗传学杂志》1998年第4期242-245,共4页Chinese Journal of Medical Genetics

基　　金：国家教委博士点基金

摘　　要：目的建立快速、准确、有效的Ｗｉｌｓｏｎ病（Ｗｉｌｓｏｎｄｉｓｅａｓｅ，ＷＤ）患者早期诊断及杂合子检测的基因诊断技术。方法应用Ｄ１３Ｓ３０１、Ｄ１３Ｓ３１６、Ｄ１３Ｓ２９６、ＡＦＭ２３８ｖｃ３、ＡＦＭ０８４ｘｃ５等５个微卫星ＤＮＡ（ｓｈｏｒｔｔａｎｄｅｍｒｅｐｅａｔ，ＳＴＲ），经聚合酶链反应（ＰＣＲ），扩增片段长度多态性－聚丙烯酰胺凝胶电泳（ＡＦＬＰ－ＰＡＧＥ），对２３个ＷＤ家系１２０名成员的ＤＮＡ进行单体型分析。结果２例拟诊患者得到确诊。在３１例ＷＤ同胞中，检出肯定症状前患者４例、杂合子８例、正常人１７例、可疑患者２例。结论ＳＴＲ单体型可提供高信息量的遗传诊断信息。Objective To develop a rapid,exact and effective technique of gene diagnosis for the early diagnosis of patients with Wilson's disease(WD) and detection of WD heterozygotes. Methods Five microsatellite (or short tandem repeat,STR) loci at D13S301,D13S316,D13S296,AFM238vc3 and AFM084xc5 were amplified by polymerase chain reaction(PCR).D13S301 was a locus within WD gene,the others were in the flank of WD gene.After detection of amplified fragment length polymorphism of the five STR loci with denaturing polyacrylamide gel electrophoresis(AFLPPAGE),the STR haplotypes analyses were performed on 120 members in 23 WD families. Results The five STR loci had high polymorphism information content(PIC).PICs of D13S301, D13S316,D13S296,AFM084xc5 and AFM238vc3 were 0.89,0.93,0.91,0.91 and 0.94 respectively.The STR haplotypes analyses confirmed that the two patients previously diagnosed doubtfully were WD patients definitely. Among 31 sibs without any clinical symptoms,4 were diagnosed as presymptomatic patients,8 as WD heterozygotes,17 as normal homozygotes and 2 as doubtful presymptomatic patients. Conclusion Heplotypes analysis of the five STR loci can provide high genetic information.It provides a new important technique in gene diagnosis of WD.

关 键 词：肝豆状核变性 微卫星DNA 单体型 杂合子 

分 类 号：R742.404[医药卫生—神经病学与精神病学]