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一例2N型遗传性血管性血友病家系的表型诊断和基因型分析

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  • 1.上海交通大学医学院附属瑞金医院北院检验科,上海 201801;
    2.上海交通大学医学院附属瑞金医院检验科,上海 200025

收稿日期: 2018-03-26

  网络出版日期: 2018-04-25

基金资助

国家自然基金青年科学基金(81601823); 上海市科委自然科学基金(16ZR1422100)

Phenotype and genotype analysis of a Chinese pedigree with 2N type von Willebrand disease

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  • 1. Department of Clinical Laboratory, Ruijin North Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 201801, China;
    2. Department of Clinical Laboratory, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China

Received date: 2018-03-26

  Online published: 2018-04-25

摘要

目的:探讨1个遗传性血管性血友病(von Willebrand disease,vWD)2N型家系的表型诊断和基因型分析结果,明确患者的发病机制。方法:对该家系的先证者和家系成员进行出血时间、活化部分凝血活酶时间、瑞斯托霉素诱导的血小板聚集(ristocetin induced platelet aggregation,RIPA)试验、血管性血友病因子(von Willebrand factor,vWF)瑞斯托霉素辅因子、vWF抗原、vWF活性测定、vWF胶原结合试验、凝血因子Ⅷ(coagulation factor Ⅷ,FⅧ)活性、vWF与FⅧ结合试验检测,并作出表型诊断。提取先证者的外周血基因组DNA,用PCR法扩增VWF基因和F8基因的所有外显子及侧翼序列,通过直接测序分析VWF基因和F8基因变异。结果:vWD家系先证者的活化部分凝血活酶时间和出血时间明显延长,血浆瑞RIPA试验、vWF瑞斯托霉素辅因子、vWF抗原、vWF活性和vWF胶原结合试验检测结果均正常,FⅧ活性下降,vWF与FⅧ的结合能力降低。对先证者进行基因测序,发现其VWF基因19号外显子存在c.2446C>T(p.Arg816Trp)错义突变,其儿子在该位点为杂合突变,而先证者及家系成员的F8基因未发现突变。结论:c.2446C>T(p.Arg816Trp)错义突变是导致该家系先证者发生2N型遗传性vWD的原因。

本文引用格式

金佩佩, 梁茜, 戴菁, 丁秋兰, 孙顺昌, 王学锋 . 一例2N型遗传性血管性血友病家系的表型诊断和基因型分析[J]. 诊断学理论与实践, 2018 , 17(02) : 151 -154 . DOI: 10.16150/j.1671-2870.2018.02.006

Abstract

Objective: To analyze the phenotype and genotype of one Chinese pedigrees with von Willebrand disease and to investigate the molecular pathogenesis of the disease. Methods: Indices including bleeding time(BT), activated partial thromboplastin time (APTT), ristocetin-induced platelet aggregation(RIPA), von Willebarand factor-ristocetin cofactor (vWF:Rco), von Willebrand factor antigen (vWF:Ag), von Willebrand factor activity (vWF:Act), von Willebrand factor collagen binding assay (vWF:CB) and von Willebrand factor FⅧ binding assay (vWF: FⅧ:B) were detected for phenotype diagnosis. Peripheral blood DNA was extracted, and all of the exons and exon-intron boundaries of the VWF and F8 gene were amplified by polymerase chain reaction (PCR) and analyzed with direct sequencing. Results: The results revealed that APTT and BT of proband were prolonged while plasma RIPA, vWF:Rco, vWF:Ag, vWF:Act and vWF:CB were normal. FⅧ:C and vWF: FⅧ:B were significantly decreased. Homozygous missense mutation c.2446C>T (p.Arg816Trp) in exon 19 of VWF gene was identified in proband and heterozygous mutation was identified in his son. No mutation in F8 gene was found in proband. Conclusions: Homozygous missense mutation c.2446C>T (p.Arg816Trp) in exon 19 of VWF gene is the cause of 2N type von Willebrand disease in the proband.

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