Molecular genetic diagnosis in the group of hemophilia A patients in Belarus: 12 new allelic variants in the <i>F8</i> gene

Q4 Medicine
A. V. Liubushkin, I. E. Guryanova, E. V. Dmitriev, V. R. Vertelko, E. A. Polyakova, L. I. Volkova, O. V. Aleinikova
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引用次数: 1

Abstract

Hemophilia A is the most common severe bleeding disorder caused by various genetic changes in the F8 gene, leading to coagulation factor VIII deficiency. Hemophilia A is characterized by high heterogeneity of genetic defects. The severity of hemophilia A varies depending on the type of genetic defects in the F8 gene. More than 3000 unique variants of the F8 gene are associated with the hemophilia A. Approximately 30% of genetic defects occur de novo. The aim of this study is to determine the spectrum of genetic defects in the F8 gene in children with hemophilia A in Belarus. The study was approved by the Independent Ethics Committee and the Scientific Council of the Belarusian Research Center for Pediatric Oncology, Hematology and Immunology (the Republic of Belarus). The study included 98 patients with hemophilia A, who had been treated or followed up at the Belarusian Research Center for Pediatric Oncology, Hematology and Immunology (the Republic of Belarus). Patients were categorized into 3 groups based on the severity of their disease: severe (n = 82), moderate (n = 3), and mild (n = 13). Twenty (20.4%) patients had a history of inhibitors to factor VIII. For our study, we used venous blood samples. Genomic DNA was isolated from leukocyte suspension (obtained from the whole blood samples) using phenol-chloroform extraction. All severe hemophilia A patients were prescreened for intron 22 and 1 inversions in the F8 gene using inverse and multiplex polymerase chain reaction assays, respectively. Sequencing of F8 coding regions was carried out by next generation sequencing. All clinically relevant variants were confirmed by Sanger sequencing. Genetic testing revealed that 99% of the patients with hemophilia A (n = 97) had pathogenic variants in the F8 gene. Intron 22 and intron 1 inversion mutations within the F8 gene were detected in 45.1% (n = 37) and 1.2% (n = 1) patients with severe hemophilia A, respectively. Two patients had an abnormal pattern of intron 1 inversion, not previously described in the literature. A total of 48 different variants in the F8 gene were detected in 57 patients using next generation sequencing. Eleven of the 48 genetic variants identified have not been previously reported.
白俄罗斯A型血友病患者的分子遗传学诊断:12个新等位基因变异<i>F8</i>基因
血友病A是最常见的严重出血性疾病,由F8基因的各种遗传变化引起凝血因子VIII缺乏。血友病A的特点是遗传缺陷的高度异质性。A型血友病的严重程度取决于F8基因的遗传缺陷类型。超过3000种独特的F8基因变异与a型血友病有关,大约30%的遗传缺陷是从头开始的。本研究的目的是确定白俄罗斯A型血友病儿童F8基因的遗传缺陷谱。该研究得到了白俄罗斯儿科肿瘤学、血液学和免疫学研究中心(白俄罗斯共和国)独立伦理委员会和科学委员会的批准。该研究包括98名A型血友病患者,他们在白俄罗斯儿科肿瘤学、血液学和免疫学研究中心(白俄罗斯共和国)接受过治疗或随访。根据病情严重程度将患者分为3组:重度(n = 82)、中度(n = 3)和轻度(n = 13)。20例(20.4%)患者有因子VIII抑制剂史。在我们的研究中,我们使用静脉血样本。基因组DNA用苯酚-氯仿萃取法从白细胞悬浮液(从全血样本中获得)中分离出来。所有严重A型血友病患者分别采用逆反应和多重聚合酶链反应法对F8基因内含子22和1反转进行预筛选。F8编码区测序采用下一代测序。所有临床相关变异均通过Sanger测序证实。基因检测显示99%的A型血友病患者(n = 97)存在F8基因致病性变异。45.1% (n = 37)和1.2% (n = 1)的重度A型血友病患者分别检出F8基因22和1内含子倒置突变。两名患者有异常模式的内含子1倒置,以前没有在文献中描述。使用下一代测序技术,在57例患者中共检测到48种不同的F8基因变异。发现的48种基因变异中有11种以前没有报道过。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Pediatric Hematology/Oncology and Immunopathology
Pediatric Hematology/Oncology and Immunopathology Medicine-Pediatrics, Perinatology and Child Health
CiteScore
0.40
自引率
0.00%
发文量
49
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