Medical University Innsbruck, Anichstr 35, A-6020 Innsbruck, Austria.
Am J Clin Pathol. 2010 Feb;133(2):322-30. doi: 10.1309/AJCPZSBTD2BWOMVL.
Analysis of von Willebrand factor (vWF) multimers allows classification of the subtypes of von Willebrand disease (vWD) in human serum and platelet lysates. A novel method for multimer analysis of vWF by 2-chamber, vertical (sodium dodecyl sulfate), agarose gel electrophoresis, designed for comparing discontinuous high- and low-resolving gels for plasma and platelets, followed by Western blotting and high-sensitivity fluorescence detection (HSFD) of cyanine (Cy)5-labeled vWF multimers is presented. HSFD shows that this method has high discriminatory power for visualization and densitometric analysis of platelets and plasma vWF multimers in various types of vWD and allows rapid classification of vWD types, to separate types 2A and 2B. The described procedures of vWF multimer analysis with high-sensitivity Cy5 fluorescence detection and direct comparison of high- and low-resolving gels for screening and detection of the complete range of high- and low-molecular vWF multimers is efficient and useful for screening, detecting, and classifying vWD subtypes and makes this method diagnostically and clinically relevant.
分析血管性血友病因子 (vWF) 多聚体可将人血清和血小板裂解物中的血管性血友病 (vWD) 亚型进行分类。本研究设计了一种用于 2 室、垂直 (十二烷基硫酸钠)、琼脂糖凝胶电泳分析 vWF 多聚体的新方法，该方法用于比较用于血浆和血小板的不连续高分辨和低分辨凝胶，并通过 Cy5 标记的 vWF 多聚体的 Western 印迹和高灵敏度荧光检测 (HSFD) 进行分析。HSFD 表明，该方法对于可视化和密度计分析各种类型 vWD 中的血小板和血浆 vWF 多聚体具有高分辨力，并允许快速分类 vWD 类型，以区分 2A 和 2B 型。该文描述的用高灵敏度 Cy5 荧光检测进行 vWF 多聚体分析的程序，并直接比较高分辨和低分辨凝胶，用于筛选和检测完整范围的高分子量和低分子量 vWF 多聚体，是一种高效且有用的方法，可用于筛选、检测和分类 vWD 亚型，并使该方法具有诊断和临床意义。
