Borràs Nina, Comes Natàlia, Ramírez Lorena, Parra Rafael, Altisent Carmen, Lorenzo-Vizcaya Álvaro, Marzo-Alonso Cristina, López-Fernández Maria-Fernanda, Canaro Mariana, Falcón-Rodríguez María, Cortes-Vidal Ángela, Rios de Paz Mario A, Rodríguez-García José Antonio, Moreto-Quintana Ana, Bandini Perla, Hobeich Carlos, Corrales Irene, Vidal Francisco
Laboratori Coagulopaties Congènites, Banc de Sang i Teixits, Barcelona, Spain.
Medicina Transfusional, Vall d'Hebron Institut de Recerca, Universitat Autònoma de Barcelona (VHIR-UAB), Barcelona, Spain.
Haemophilia. 2025 Jul;31(4):734-742. doi: 10.1111/hae.70075. Epub 2025 Jun 27.
Inherited coagulation factor deficiencies (ICFD) result from plasma protein deficiencies, impacting blood coagulation cascade and leading to haemorrhagic diathesis. Advancements in next-generation sequencing (NGS) technology have enabled high-throughput methods for molecular ICFD diagnosis. However, detailed descriptions of clinical applications and routine laboratory experiences in this field remain scarce.
This study presents the results from a real-world experience using an NGS-based gene panel for routine molecular diagnosis, applied to more than 500 ICFD patients in Spain.
A custom NGS gene panel targeting 22 ICFD-related genes was validated using 20 patients with known variants. Subsequently, the panel was applied to 515 ICFD patients from 28 Spanish hospitals. Structural variants were detected by multiplex ligation-dependent probe amplification.
Among the 515 patients analysed, 402 had complete phenotypic data specified in the genetic study form, 83 had incomplete data, and 30 were potential haemophilia carriers. Identification disease-causing variant rates were 69%, 58% and 53%, respectively. Candidate variants were identified in 74% of cases. A total of 460 variants across 18 genes were identified, 302 unique variants, with 37% being novel disease-causing variants.
This study represents the largest analysis of ICFD patients conducted in Spain, providing significant insights into the molecular epidemiology of these disorders. It underscores the critical role of NGS in routine clinical practice while addressing challenges faced by genetic laboratories. The findings highlight a growing shift among haematologists towards integrating genetic studies early in diagnostic workflows alongside phenotypic assessments to enhance the accuracy and efficiency of ICFD diagnosis.
遗传性凝血因子缺乏症（ICFD）是由血浆蛋白缺乏引起的，影响血液凝固级联反应并导致出血素质。下一代测序（NGS）技术的进步使得分子ICFD诊断的高通量方法成为可能。然而，该领域临床应用和常规实验室经验的详细描述仍然很少。
本研究展示了使用基于NGS的基因panel进行常规分子诊断的真实世界经验结果，该基因panel应用于西班牙500多名ICFD患者。
使用20名已知变异的患者对靶向22个ICFD相关基因的定制NGS基因panel进行验证。随后，该基因panel应用于来自28家西班牙医院的515名ICFD患者。通过多重连接依赖探针扩增检测结构变异。
在分析的515名患者中，402名在基因研究表格中有完整的表型数据，83名数据不完整，30名是潜在的血友病携带者。致病变异的识别率分别为69%、58%和53%。在74%的病例中鉴定出候选变异。共鉴定出18个基因的460个变异，其中302个是独特变异，37%是新的致病变异。
本研究是西班牙对ICFD患者进行的最大规模分析，为这些疾病的分子流行病学提供了重要见解。它强调了NGS在常规临床实践中的关键作用，同时解决了基因实验室面临的挑战。研究结果突出了血液学家越来越倾向于在诊断工作流程中尽早将基因研究与表型评估相结合，以提高ICFD诊断的准确性和效率。
