학술논문
一个Gly363Ser突变导致遗传性凝血因子Ⅹ缺乏症家系的表型及基因型分析 / Genotypic and phenotypic analysis of a case with inherited coagulation factor Ⅹ deficiency
Document Type
Academic Journal
Author
陈陶; 李帆帆; 舒旷怡; 柳洁; 沈晨芳; 章赵华; 金速速; 王晓欧; 江明华; Chen Tao; Li Fanfan; Shu Kuangyi; Liu Jie; Shen Chenfang; Zhang Zhaohua; Jin Susu; Wang Xiaoou; Jiang Minghua
Source
中华医学遗传学杂志 / Chinese Journal of Medical Genetics. 35(4):544-547
Subject
Language
Chinese
ISSN
1003-9406
Abstract
目的 分析1例凝血因子Ⅹ (coagulable factor Ⅹ,FⅩ)缺陷症家系的表型及基因型,并探讨F10基因突变与表型的关系.方法 用一期凝固法测定先证者及其家系成员的凝血酶原时间(prothrombin time,PT)、活化部分凝血酶时间(activated partial thromboplastin time,APTT)、纤维蛋白(fibrinogen,FIB)及血浆FⅡ活性(FⅡactivity,FⅡ∶C)、血浆FⅦ活性(FⅦactivity,FⅦ∶C)、血浆FⅨ活性(FⅨactivity,FⅨ∶C)、血浆FⅩ活性(FⅩ activity,FⅩ∶C)等指标进行表型诊断;用ELISA法检测FⅩ抗原含量(FⅩ antigen,FⅩ∶Ag);用PCR对先证者及其家系成员F10基因8个外显子及其侧翼序列、5'和3'非翻译区进行扩增,产物纯化后直接进行正向和反向测序;同时对100名健康对照DNA相应突变区域测序以排除基因多态性;应用软件分析突变位点序列和蛋白质构象改变特点及同源比对分析.结果 先证者PT、APTT、FⅩ∶C、FⅩ∶Ag均有异常,分别为16.1s、49.0 s、27％、56％,其他凝血表型指标均正常;先证者母亲PT、APTT、FⅩ∶C、FⅩ∶Ag分别为14.8s、37.4s、44％、34％;先证者外祖母PT、APTT、FⅩ∶C、FⅩ∶Ag分别为15.8 s、42.2 s、31％、45％;父亲的凝血指标均正常.先证者、母亲和外祖母均为F10第8外显子g.28076G＞A杂合突变,导致p.Gly363Ser,父亲无此突变.p.Gly363Ser突变导致FⅩ蛋白二级结构和三维空间结构的改变,导致活性降低.结论 该先证者的家系中存在F10遗传性杂合突变g.28076G＞A(p.Gly363Ser),且该突变与FⅩ水平降低有关,为国内尚未见报道的突变.
Objective To explore the correlation between F10 gene mutation and its phenotype in a Chinese pedigree affected with F Ⅹ deficiency.Methods Prothrombin time (PT),activated partial thromboplastin time(APTT),fibrinogen,F Ⅱ activity(F [Ⅱ ∶ C),F Ⅶ activity (F Ⅶ ∶ C),F Ⅸ activity (FⅨ ∶ C),FⅩ activity(FⅩ ∶ C) were determined with a one-stage clotting assay.The FⅩ antigen(FⅩ ∶ Ag) was detected with an enzyme linked immunosorbent assay(ELISA).The 8 exons,introns and 5'and 3'untranslated regions(UTR) of the F10 gene of the proband and her family members were subjected to PCR amplification and Sanger sequencing.Suspected mutation was confirmed by reverse sequencing.Polymorphisms were excluded by direct sequencing of 100 healthy individuals.Results The PT and APTT of the proband have prolonged to 16.1 s and 49.0 s,respectively.Her FⅩ ∶ C and FⅩ ∶ Ag were reduced by 27％ and 56％,and her mother's PT,APTT,FⅩ ∶C and FⅩ ∶Ag were 14.8 s,37.4 s,44％,34％,respectively.Her grandmother's PT,APTT,FⅩ ∶ C and FⅩ ∶ Ag were 15.8 s,42.2 s,31％,45％,respectively.The results of her father and other family members were all within the normal range.Genetic analysis has revealed a heterozygous G＞A mutation in the proband at position 28076 in exon 8 of the F10 gene,which resulted in a p.Gly363Ser substitution.The same mutation was also found in her mother and grandmother.No mutation of the F10 gene was found in her father.Gly363Ser may result in changes in the secondary structure of the F Ⅹ protein and reduction of its activity.Conclusion The g.28076G＞ A (p.Gly363Ser) mutation of the F10 gene probably underlies the F Ⅹ deficiency in this pedigree.The mutation was discovered for the first time in Chinese patients.
Objective To explore the correlation between F10 gene mutation and its phenotype in a Chinese pedigree affected with F Ⅹ deficiency.Methods Prothrombin time (PT),activated partial thromboplastin time(APTT),fibrinogen,F Ⅱ activity(F [Ⅱ ∶ C),F Ⅶ activity (F Ⅶ ∶ C),F Ⅸ activity (FⅨ ∶ C),FⅩ activity(FⅩ ∶ C) were determined with a one-stage clotting assay.The FⅩ antigen(FⅩ ∶ Ag) was detected with an enzyme linked immunosorbent assay(ELISA).The 8 exons,introns and 5'and 3'untranslated regions(UTR) of the F10 gene of the proband and her family members were subjected to PCR amplification and Sanger sequencing.Suspected mutation was confirmed by reverse sequencing.Polymorphisms were excluded by direct sequencing of 100 healthy individuals.Results The PT and APTT of the proband have prolonged to 16.1 s and 49.0 s,respectively.Her FⅩ ∶ C and FⅩ ∶ Ag were reduced by 27％ and 56％,and her mother's PT,APTT,FⅩ ∶C and FⅩ ∶Ag were 14.8 s,37.4 s,44％,34％,respectively.Her grandmother's PT,APTT,FⅩ ∶ C and FⅩ ∶ Ag were 15.8 s,42.2 s,31％,45％,respectively.The results of her father and other family members were all within the normal range.Genetic analysis has revealed a heterozygous G＞A mutation in the proband at position 28076 in exon 8 of the F10 gene,which resulted in a p.Gly363Ser substitution.The same mutation was also found in her mother and grandmother.No mutation of the F10 gene was found in her father.Gly363Ser may result in changes in the secondary structure of the F Ⅹ protein and reduction of its activity.Conclusion The g.28076G＞ A (p.Gly363Ser) mutation of the F10 gene probably underlies the F Ⅹ deficiency in this pedigree.The mutation was discovered for the first time in Chinese patients.