两个遗传性凝血因子Ⅻ缺陷症家系的表型及基因分析  

Phenotypic and genetic analysis of two pedigrees affected with hereditary coagulation FⅫ deficiency

作　　者：李姗姗[1] 沈晨芳 舒旷怡[1] 柳洁[1] 王晓欧[1] 李帆帆[1] 杨啸 章赵华[1] 陈碧[1] 江明华[1] Li Shanshan;Shen Chenfang;Shu Kuangyi;Liu Jie;Wang Xiaoou;Li Fan fan;Yang Xiao;Zhang Zhaohua;Chen Bi;Jiang Minghua(Department of Laboratory Medicine,the Second Affiliated Hospital,Wenzhou Medical University,Wenzhou,Zhejiang 325027,China)

机构地区：[1]浙江省温州医科大学附属第二医院检验科,325027

出　　处：《中华医学遗传学杂志》2018年第6期800-803,共4页Chinese Journal of Medical Genetics

基　　金：浙江省自然科学基金(LY13H200003);温州市公益性科技计划(Y20140674);浙江省医药卫生科技项目(2016RCA023,2015KYA155).

摘　　要：目的分析两个遗传性凝血因子Ⅻ(coagulation factor Ⅻ,FⅫ)缺陷症家系的表型及基因突变,探讨其分子发病机制。方法一期凝固法检测血浆凝血酶原时间,活化部分凝血活酶时间(activatedpartial thromboplastin time,APTT),纤维蛋白原,凝血酶时间,凝血因子Ⅷ、Ⅸ、Ⅺ、Ⅻ的促凝活性(FⅧ:C、FIX:C、FⅪ:C、FⅫ:C);ELISA法检测FⅫ抗原(FⅫ:Ag);DNA测序分析患者F/2基因的14个外显子及其侧翼序列;MegAlign、PYMOL等平台或软件分析基因突变位点物种保守性和蛋白质构象改变。结果两家系先证者均表现为APTT明显延长,FⅫ:C、FⅫ:Ag明显降低,分别为126.3s、2%、10%和122.0S、1%、11%。对两个家系的成员进行基因分析,共发现4种突变,其中3种为未报道的突变,包括第5内含子3’端的g.5972G>A(IVS5-1G>A)剪接突变、第14外显子g.8810_8814del GTCTA的小片段缺失、以及第7外显子g.6259G>A(p.Pro182Leu)。此外IVS13-1G>A为已报道突变。结论两个家系成员中发现4种基因突变,可能与这两个家系FⅫ缺陷有关,其中3种(g.5972G>A、g.8810_8814 del GTCTA及g.6259G>A)为尚未见报道的突变。Objective To carry out phenotypic and genotypic analysis for two Chinese pedigrees affected with coagulation factor Ⅻ(FⅫ)deficiency.Methods Plasma prothrombin time (PT),activated partial thromboplastin time (APTT),fibrinogen (FIB),thrombin time (TT),and blood coagulation factor Ⅷ,Ⅸ,Ⅺ,Ⅻ activity (FⅧ:C,FⅨ:C,FⅪ:C,FⅫ:C)were determined with one stage clotting assay on a STAGO coagulation analyzer.FⅫ antigen was determined with an enzyme linked immunosorbent assay (ELISA).The 14 exons and their flanking sequences of the F12 gene were subjected to PCR amplification and Sanger sequencing.The conservation and structure of mutant protein were analyzed with MegAlign software and PYMOL software.Results The APTT of the probands was significantly prolonged,while their FⅫ:C and FⅫ:Ag were significantly reduced.Genetic analysis of the proband has revealed three novel mutations in the F12 gene,including g.5972G>A splice site mutation in intron 5, g.8810_8814delGTCTA in exon 14,and g.6259G>A (p.Pro182Leu)in exon 7.In addition,a previously known mutation IVS13-1G>A has been found.Conclusion Four mutations have been identified in the two Chinese pedigrees,among which three were novel.Above mutations probably played a role in the defect of FⅫ in the two pedigrees.

关 键 词：凝血因子Ⅻ缺乏症 杂合子 突变 

分 类 号：R596[医药卫生—内科学]