vWD家系基因连锁分析的应用研究

Clinical Application of Linkage Analysis for vWD Family

目的 探讨 v WF基因内含子 4 0中 nt1890 - 1990和 nt2 2 15 - 2 380两个四核苷酸 STR位点的扩增长度片段多态性 (am p- FL P)在血管性血友病 (v WD)基因诊断和遗传咨询中的应用。方法 对 8个家系中的相关成员v WF基因内含子 4 0中两个 STR位点进行聚合酶链反应 (PCR)扩增 ,采用聚丙烯酰胺凝胶电泳 (PAGE)与银染技术对 PCR产物进行检测 ,通过两位点的 am p- FL P进行基因连锁分析。结果 8个家系 v WF基因内含子 4 0中nt1890 - 1990位点和 nt2 2 15 - 2 380位点各检出 5种等位基因 ,均有效确定出与致病基因连锁的单倍型组成 ,结合临床症状 ,为遗传咨询提供了线索。结论 v WF基因内含子 4 0的两个四核苷酸重复位点是进行家系间接基因诊断较好的遗传标记 ;PCR- PAGE能方便、快速与准确地检测 v WF基因内含子 4 0的两个 STR位点的片段长度多态性 ,适用于 v WD的家系分析和遗传咨询。

Objective To explore the application of amplified fragment length polymorphism (amp-FLP) of short tandem repeat (STR) within intron 40 of vWF gene in the gene diagnosis and genetic consulting service for von Willebrand disease (vWD). Methods We isolated DNA from the blood of members in 8 families with vWD, measured the relative parameters of vWD simultaneously; and amplified the fragment length polymorphism of two loci (nt1890-1990 and nt2215-2380) within intron 40 of vWF gene using PCR. The RCR products were analyzed by means of polyacrylamide gel electrophoresis (PAGE) and silver staining. Results Five types of amp-FLP were identified on nt1890-1990 and nt2215-2380 respectively. Haplotypes could be identified to link with defective vWF gene in these families. Conclusion Combination of PCR and PAGE is a fast and practical method for carrying out family analysis of inherited disease; nt1980-1990 and nt2215-2380 of vWF gene are two ideal genetic labels in linkage analysis and hereditary consultation of vWD family.