Department of Diagnostics, Spedali Civili of Brescia, Brescia, Italy.
Electrophoresis. 2013 Mar;34(6):818-24. doi: 10.1002/elps.201200538.
Abnormalities in plasma von Willebrand factor (vWF) concentration and function result in von Willebrand disease (vWD). The diagnosis requires a battery of tests such as screening procedures, confirmatory tests, phenotypic characterization, and genotyping. The phenotypic testing (multimer pattern analysis) is important in order to subclassify the hereditary and the acquired forms of vWD. Only few laboratories are skilled to perform this analysis. The extreme range of protein size from 250 kDa monomer to over 20,000 kDa multimers requires a time-consuming procedure (3-4 days) and presents many technical difficulties. To standardize the method and to overcome technical difficulties, we developed a rapid and sensitive semi-automated method to visualize the multimeric structure of vWF. The semi-automated method we present performs the electrophoresis of patient's plasma in 120 min on a precast gel. Gels are suitable for the G26 Interlab instrumentation. After gel blotting, the method allows visualization of the vWF multimer pattern directly on the membrane. We reduced the time required from 72 to 8 h and we propose this test for the first level screening of vWF multimer deficiency.
血浆血管性血友病因子 (vWF) 浓度和功能异常会导致血管性血友病 (vWD)。诊断需要一系列测试，如筛选程序、确认测试、表型特征分析和基因分型。表型测试（多聚体模式分析）对于对遗传性和获得性 vWD 进行分类很重要。只有少数实验室具备进行这种分析的能力。蛋白质大小的范围从 250kDa 单体到超过 20000kDa 的多聚体非常大，需要耗时的程序（3-4 天）并存在许多技术困难。为了标准化方法并克服技术困难，我们开发了一种快速灵敏的半自动化方法来可视化 vWF 的多聚体结构。我们提出的半自动方法可在预制凝胶上在 120 分钟内对患者血浆进行电泳。凝胶适用于 G26 Interlab 仪器。凝胶印迹后，该方法可直接在膜上可视化 vWF 多聚体模式。我们将所需时间从 72 小时缩短到 8 小时，并建议将该测试用于 vWF 多聚体缺乏症的一级筛查。
