多重聚合酶链反应检测vWF基因内微卫星DNA与vWD患者家系分析  

Linkage analysis of vWD family by multiPCR assay of microsatellite DNA in vWF gene

作　　者：杜玲珍[1,2] 于立志[1,2] 王鸿利[1,2] 储海燕[1,2] 诸江[1,2] 王振义[1,2] 

机构地区：[1]上海第二医科大学瑞金医院儿内科 [2]上海血液学研究所

出　　处：《中华血液学杂志》1998年第3期118-121,共4页Chinese Journal of Hematology

基　　金：卫生部科学研究基金;胡应洲基金

摘　　要：目的：研究中国人ｖＷＦ基因内含子４０中２个微卫星位点扩增片段长度多态性（ａｍｐ－ＦＬＰ），并进行血管性血友病（ｖＷＤ）家系分析。方法：多重聚合酶链反应（ＰＣＲ）一次反应扩增Ａ、Ｂ两个片段，用变性聚丙烯酰胺凝胶电泳和高敏感度银染方法分析Ａ、Ｂ位点的ａｍｐ－ＦＬＰ。对１１２名上海地区正常人和两个ｖＷＤ家系成员ＤＮＡ标本进行了检测。结果：①所检测上海人群中在Ａ位点存在７种ａｍｐ－ＦＬＰ，杂合率为７５％；Ｂ位点存在５种ａｍｐ－ＦＬＰ，杂合率为７４％。②在两个ｖＷＤ家系中发现单倍型Ａ２／Ｂ３和Ａ４／Ｂ３分别与致病基因连锁，结合临床症状，为遗传咨询提供了线索。结论：①多重ＰＣＲ检测微卫星ＤＮＡ方便、快捷，适用于临床应用进行遗传性疾病的家系分析和遗传咨询。②ｎｔ１８９０－１９９０和ｎｔ２２１５－２３８０是用于ｖＷＤ家系连锁分析和遗传咨询较好的ｖＷＦ基因内遗传标记。Objective:To set up a convenient method for analysis of the amplified fragment length polymorphism(ampFLP) of two microsatellite DNA in vWF gene.Methods:MultiPCR followed by denatured PAGE and silver stain were used to analyze the ampFLP of two locus (A and B) on one gel.DNA samples from 112 normal people and two vWD families in the Shanghai area were assayed.Results:① seven and 5 types of ampFLP were identified on loci A and B, respectively.The heterozygote rates were 75% and 74%,respectively.②Haplotypes A2/B3 and A4/B3 were found to link with defective vWF genes in the two vWD families,respectively.Conclusion:①MultiPCR is a fast and practical method to carry out family analysis of inherited diseases.②nt19801990 and nt22152380 of vWF gene are two ideal genetic labels in linkage study and hereditary consultation of vWD family.

关 键 词：血管性血友病 血友病 聚合酶链反应 αmp-FLP 

分 类 号：R554.102[医药卫生—血液循环系统疾病]