Habart D, Smejkal P, Matýsková M, Turek P, Hrachovinová I, Vorlová Z
Ustav hematologie a krevní transfuze, Praha.
Cas Lek Cesk. 2003;142(6):373-6.
von Willebrand disease is an inherited bleeding disorders caused by mutations in the von Willebrand factor gene. We attempted to characterise the phenotype and the genotype in the first five families in Czech Republic affected by this heterogeneous disorder.
The level of FVIII was measured by the one stage assay, the vWF:Ag by the immunoelectrophoresis, vWF:RiCo by aggregometry. For the vWF multimer analysis a western blot based technique was used. The vWF binding to FVIII was evaluated by the ELISA method. Two families were classified as the type 2A, one as the type 2B and two as the combined type 1/2N. Based on that knowledge, parts of the vWF gene were selected for genetic analysis. The previously described mutations Arg1374His and Gly1579Arg were identified in two families with the type 2A. In the family with type 2B a substitution Arg1308Cys was detected. In one family with the type 1/2N, two different previously described defects were found on the separate alleles of the vWF gene: a deletion of cytosine 2435 and a polymorphism Arg854Gln. Compound heterzygotes had the type 1/2N phenotype, while a carriers of the deletion had type 1 phenotype. In the second type 1/2N family, only the amino acid substitutions Thr791Me was found explaining the qualitative defect. A mutation underlying the quantitative deficiency needs to be searched for throughout the entire vWF gene.
Based on the characterisation of the phenotype and genotype, five apparently unrelated families with the von Willebrand disease were diagnosed according to the revised classification. Our work represents laboratory basis for further studies into von Willebrand disease in Czech Republic.
血管性血友病是一种由血管性血友病因子基因突变引起的遗传性出血性疾病。我们试图对捷克共和国受这种异质性疾病影响的前五例家族的表型和基因型进行特征分析。
采用一期法检测FVIII水平，免疫电泳法检测vWF:Ag，凝集法检测vWF:RiCo。vWF多聚体分析采用基于蛋白质印迹的技术。通过ELISA法评估vWF与FVIII的结合。两个家族被分类为2A型，一个为2B型，两个为1/2N复合型。基于这些认识，选择vWF基因的部分区域进行基因分析。在两个2A型家族中鉴定出先前描述的突变Arg1374His和Gly1579Arg。在2B型家族中检测到替代突变Arg1308Cys。在一个1/2N型家族中，在vWF基因的不同等位基因上发现了两个不同的先前描述的缺陷：胞嘧啶2435缺失和多态性Arg854Gln。复合杂合子具有1/2N型表型，而缺失携带者具有1型表型。在第二个1/2N型家族中，仅发现氨基酸替代Thr791Met解释了定性缺陷。需要在整个vWF基因中寻找导致定量缺陷的突变。
基于表型和基因型的特征分析，根据修订后的分类法诊断了五个明显无关的血管性血友病家族。我们的工作为捷克共和国进一步研究血管性血友病提供了实验室依据。
