一个遗传性抗凝血酶与凝血因子Ⅶ联合缺陷症家系的临床特征和基因分析  

Clinical characteristics and genetic analysis of a combined inherited antithrombin and factorⅦdeficiency pedigree

作　　者：张海月 王明山[1] 谢海啸[1] 刘斯奇 罗莎莎 杨丽红[1] 周星星[1] 金艳慧[1] Zhang Haiyue;Wang Mingshan;Xie Haixiao;Liu Siqi;Luo Shasha;Yang Lihong;Zhou Xingxing;Jin Yanhui(Department of Clinical Laboratory,the First Affiliated Hospital of Wenzhou Medical University,Wenzhou 325015,China)

机构地区：[1]温州医科大学附属第一医院医学检验中心,温州325015

出　　处：《中华检验医学杂志》2020年第6期635-639,共5页Chinese Journal of Laboratory Medicine

基　　金：浙江省科技厅公益技术研究计划基金(LGF18H080003);浙江医药卫生科技项目(2017KY457)。

摘　　要：目的对一个遗传性抗凝血酶(AT)与凝血因子Ⅶ(FⅦ)联合缺陷症家系进行临床特征和基因突变分析,探讨AT基因和F7基因突变与疾病发生的关系。方法家系调查。收集2018年11月来温州医科大学附属第一医院就诊的先证者及其家系成员血液和临床资料(共3代16人),检测凝血酶原时间(PT)、活化部分凝血活酶时间(APTT)、纤维蛋白原(FIB)、抗凝血酶活性(AT:A)、抗凝血酶抗原(AT:Ag)、蛋白C活性(PC:A)、蛋白S活性(PS:A)、FⅦ活性(FⅦ:C)及FⅦ抗原(FⅦ:Ag)等指标以明确诊断。提取先证者及其家系成员外周血基因组DNA,用DNA直接测序法分析AT基因和F7基因全部外显子、侧翼及5′、3′非编码区,寻找基因异常位点,并通过克隆测序及反向测序进行验证。结果先证者(Ⅱ6)AT:A和AT:Ag明显下降,分别为46%和135 mg/L(参考范围为250~360 mg/L);其家庭部分成员(父亲Ⅰ2、小姑Ⅰ4、堂姐Ⅱ1、表姐Ⅱ3、小弟弟Ⅱ8、侄子Ⅲ3)AT:A和AT:Ag均降低至正常人的50%左右;其父亲(Ⅰ2)、小姑(Ⅰ4)、大弟弟(Ⅱ7)、小弟弟(Ⅱ8)、侄子(Ⅲ3)FⅦ:C分别为45%、50%、48%、47%和48%,而FⅦ:Ag正常。基因分析显示先证者(Ⅱ6)及其家庭部分成员(父亲Ⅰ2、小姑Ⅰ4、堂姐Ⅱ1、表姐Ⅱ3、小弟弟Ⅱ7、侄子Ⅲ3)AT基因5′非编码区存在rs3138521多态性;其父亲(Ⅰ2)、小姑(Ⅰ4)、大弟弟(Ⅱ7)、小弟弟(Ⅱ8)、侄子(Ⅲ3)均存在F7基因8号外显子c.1091G>A杂合错义突变,导致p.Arg304Gln。结论抗凝血酶与凝血因子Ⅶ分别存在rs3138521多态性和c.1091G>A杂合错义突变,可能是导致该家系成员AT与FⅦ联合缺陷的分子机制。Objective To study the clinical characteristics and gene mutations in a family with combined inherited antithrombin(AT)and factorⅦ(FⅦ)deficiency,and explore the relationship between AT gene,F7 gene mutations and diseases.Methods Pedigree investigation.Blood samplesand clinical dataswere collected fromthe proband and her family members(a total of 16 people in 3 generations)who admitted to the First Affiliated Hospital of Wenzhou Medical University in November 2018.The prothrombin time(PT),activated partial thromboplastin time(APTT),fibrinogen(FIB),antithrombin activity(AT:A),antithrombin antigen(AT:Ag),protein C activity(PC:A),protein S activity(PS:A),FⅦactivity(FⅦ:C),FⅦantigen(FⅦ:Ag)and other indicators were detectedto confirm the diagnosis.DNA direct sequencing analysis of all exons,flanking sequences,5′and 3′untranslated regions of AT genes and F7 genes,and the mutation sites were confirmed by clone sequencingor reverse sequencing.Results The AT:A and AT:Ag of the proband were 46%and 135 mg/L,respectively(reference range:250-360 mg/L),some of her family members′s(father,aunt,two cousins,younger brother and nephew)AT:A and AT:Ag were reduced to 50%of normal range.Her father(Ⅰ2),aunt(Ⅰ4),elder brother(Ⅱ7),younger brother(Ⅱ8),and nephew(Ⅲ3)′s FⅦ:C were 45%,50%,48%,47%and 48%,respectively;and their FⅦ:Ag was within the normal range.Genetic analysis revealed that the proband(Ⅱ6)and some of her family members(father,aunt,two cousins,younger brother and nephew)took rs3138521 polymorphism in the 5′untranslated region of AT gene.Her father(Ⅰ2),aunt(Ⅰ4),elder brother(Ⅱ7),younger brother(Ⅱ8),nephew(Ⅲ3)took c.1091G>A heterozygous missense mutationin exon 8 of F7 gene,resulting in p.Arg304Gln.Conclusion The rs3138521 in AT gene and c.1091G>A in F7 gene,which may be the molecular mechanism leading to combined inherited AT and FⅦdeficiency in this family.

关 键 词：抗凝血酶缺陷症 凝血因子Ⅶ缺陷症 基因突变 临床特征 

分 类 号：R596.1[医药卫生—内科学]