Fidalgo Teresa, Salvado Ramon, Corrales Irene, Pinto Silva Catarina, Borràs Nina, Oliveira Ana, Martinho Patricia, Ferreira Gisela, Almeida Helena, Oliveira Cristina, Marques Dalila, Gonçalves Elsa, Diniz MJoão, Antunes Margarida, Tavares Alice, Caetano Gonçalo, Kjöllerström Paula, Maia Raquel, Sevivas Teresa S, Vidal Francisco, Ribeiro Leticia
Teresa Fidalgo, Centro Hospitalar e Universitário de Coimbra (CHUC), Serviço de Hematologia Clínica, Unidade de Trombose e Hemostase, Av Afonso Romão Coimbra 3000-602, Portugal, Tel.: +351 239 480 370, Fax: +351 239 717 216, E-mail:
Thromb Haemost. 2016 Jul 4;116(1):17-31. doi: 10.1160/TH15-07-0604. Epub 2016 Mar 17.
The diagnosis of von Willebrand disease (VWD), the most common inherited bleeding disorder, is characterised by a variable bleeding tendency and heterogeneous laboratory phenotype. The sequencing of the entire VWF coding region has not yet become a routine practice in diagnostic laboratories owing to its high costs. Nevertheless, next-generation sequencing (NGS) has emerged as an alternative to overcome this limitation. We aimed to determine the correlation of genotype and phenotype in 92 Portuguese individuals from 60 unrelated families with VWD; therefore, we directly sequenced VWF. We compared the classical Sanger sequencing approach and NGS to assess the value-added effect on the analysis of the mutation distribution in different types of VWD. Sixty-two different VWF mutations were identified, 27 of which had not been previously described. NGS detected 26 additional mutations, contributing to a broad overview of the mutant alleles present in each VWD type. Twenty-nine probands (48.3 %) had two or more mutations; in addition, mutations with pleiotropic effects were detected, and NGS allowed an appropriate classification for seven of them. Furthermore, the differential diagnosis between VWD 2B and platelet type VWD (n = 1), Bernard-Soulier syndrome and VWD 2B (n = 1), and mild haemophilia A and VWD 2N (n = 2) was possible. NGS provided an efficient laboratory workflow for analysing VWF. These findings in our cohort of Portuguese patients support the proposal that improving VWD diagnosis strategies will enhance clinical and laboratory approaches, allowing to establish the most appropriate treatment for each patient.
血管性血友病（VWD）是最常见的遗传性出血性疾病，其诊断特征为出血倾向多变且实验室表型异质性。由于成本高昂，对整个VWF编码区进行测序尚未成为诊断实验室的常规操作。尽管如此，新一代测序（NGS）已成为克服这一局限性的替代方法。我们旨在确定来自60个无亲缘关系的VWD葡萄牙家庭的92名个体的基因型与表型之间的相关性；因此，我们对VWF进行了直接测序。我们比较了经典的桑格测序方法和NGS，以评估其对不同类型VWD突变分布分析的增值效应。共鉴定出62种不同的VWF突变，其中27种此前未曾报道。NGS又检测到26种额外突变，有助于全面了解每种VWD类型中存在的突变等位基因。29名先证者（48.3%）有两个或更多突变；此外，还检测到具有多效性的突变，NGS对其中7种进行了恰当分类。此外，还实现了VWD 2B与血小板型VWD（1例）、伯纳德-索利尔综合征与VWD 2B（1例）以及轻度血友病A与VWD 2N（2例）之间的鉴别诊断。NGS为分析VWF提供了高效的实验室流程。我们葡萄牙患者队列中的这些发现支持以下提议，即改进VWD诊断策略将加强临床和实验室方法，从而能够为每位患者制定最合适的治疗方案。
