机构地区:[1]温州医科大学附属第一医院医学检验中心,浙江省检验诊断及转化研究重点实验室,325015
出 处:《浙江医学》2022年第22期2432-2436,共5页Zhejiang Medical Journal
基 金:温州市基础性科研项目(Y20210111);浙江省检验诊断及转化研究重点实验室(2022E10022)。
摘 要:目的对一个Ⅱ型遗传性抗凝血酶(AT)缺陷症家系进行表型及基因突变分析,探讨该症与深静脉血栓形成的关系。方法回顾2020年2月于温州医科大学附属第一医院神经内科确诊的1例发生深静脉血栓的患者(先证者)临床资料;同时采用随机抽样法选取本院100名健康体检者作为正常对照。检测先证者及其家系成员(共3代11人)的血浆AT活性(AT:A)、AT抗原(AT∶Ag)、蛋白C活性(PC∶A)及蛋白S活性(PS∶A)等指标并进行分析。采用Sanger测序法分析SERPINC1基因全部外显子、侧翼序列、5’和3’端非编码区,寻找基因突变位点以及通过反向测序予以证实。运用3个在线生物信息学软件(MutationTaster、PolyPhen-2、LRT)分析突变氨基酸的致病性。结果先证者及其大儿子、二女儿和外孙女的AT∶A均显著降低,分别为正常对照的32%、43%、52%和48%,先证者及其家系成员PC∶A、PS∶A及AT∶Ag指标均无明显异常,表现为Ⅱ型AT缺陷症。基因分析显示:先证者SERPINC1基因第7号外显子存在c.1346T>A杂合错义突变(p.Leu417Gln),其大儿子、二女儿和外孙女也携带该突变。同源性分析表明Leu417残基在同源物种间高度保守;MutationTaster、PolyPhen-2和LRT生物信息学软件分析均显示p.Leu417Gln突变为“致病的、有害的”;蛋白模型分析显示,p.Leu417Gln突变会引起AT蛋白内部的氢键发生改变。结论该先证者及家系成员AT∶A降低与SERPINC1基因p.Leu417Gln错义突变有关;先证者出现深静脉血栓形成可能与p.Leu417Gln错义突变导致机体抗凝功能降低有关。Objective To perform pedigree analysis of a patient with type Ⅱ hereditary antithrombin(AT)deficiency causing deep vein thrombosis(DVT).Methods The clinical data of a patient diagnosed with deep vein thrombosis in the Department of Neurology,the First Affiliated Hospital of Wenzhou Medical University in February 2020 were retrospectively analyzed.A total of 100 healthy subjects who underwent physical examination were selected as normal controls.Peripheral blood samples were collected from the proband and their pedigree members(11 persons of 3 generations).The plasma AT activity(AT∶A),AT antigen(AT∶Ag),protein C activity(PC∶A)and protein S activity(PS∶A)were analyzed.All exons,flanking and 5’and 3’noncoding regions were amplified by PCR,and the PCR products were directly sequenced to search abnormal errant loci,and verified by reverse sequencing.The effect of mutations on the protein function was analyzed by bioinformatics software.Results The AT∶A was significantly reduced in the proband and its eldest son,second daughter and granddaughter at 32%,43%,52% and 48% of normal controls,respectively,while the PC∶A,PS∶A and AT∶Ag of the proband and their family member were within the normal range,presenting as type Ⅱ AT deficiency.Genetic analysis revealed the presence of in exon 7 SERPINC1 gene c.1346T>A heterozygous missense mutation(p.Leu417Gln)in the proband,his eldest son,second daughter,and granddaughter.The conservation analysis revealed that Leu417 was highly conserved among homologous species.The bioinformatics software,MutationTaster,PolyPhen-2 and LRT showed p.Leu417Gln mutation as“pathogenic,deleterious”;protein model analysis showed that p.Leu417Gln mutations cause changes in the hydrogen bonds within the AT protein.Conclusion Genetic analysis shows that the decreased AT∶A in the proband and pedigree member is associated with the missense mutations of SERPINC1 gene in p.Leu417Gln;which leads to the reduced anticoagulants and DVT in the proband.
正在载入数据...
正在载入数据...
正在载入数据...
正在载入数据...
正在载入数据...
正在载入数据...
正在载入数据...