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目的:对一个遗传性凝血酶原缺陷症家系进行凝血酶原(FII)基因突变的检测。方法:用活化部分凝血活酶时间(APTT),凝血酶原时间(PT)及FII促凝活性(FII:C)、FII抗原(FII:Ag)测定进行表型诊断;用PCR法对先证者的FII基因14个外显子及其侧翼序列和5’端非翻译区(5’UTR)、3’端非翻译区(3’UTR)序列进行扩增,PCR产物纯化后直接测序,检测其基因突变。家系成员DNA在先证者FII基因突变区域扩增后测序。突变位点经限制性内切酶分析证实。103例健康献血者作对照。结果:先证者表型诊断为凝血酶原缺陷症(Ⅰ型);FII外显子区共发现3个与文献报道的FII基因序列不同的位点,其中位于第2外显子区的为纯合突变A601G。家系分析表明先证者父亲、母亲和外祖母均为A601G杂合子。结论:纯合错义突变A601G引起的Glu29→Gly是导致本例遗传性凝血酶原缺陷症的原因。这在国际上首次报道。
Objective: To detect the mutation of prothrombin (FII) gene in a hereditary prothrombin deficiency pedigree. Methods: Phenotypic diagnosis was performed with activated partial thromboplastin time (APTT), prothrombin time (PT) and FII: C, FII antigen (FII: Ag) 14 exons of FII gene and their flanking sequences, 5 ’untranslated region (5’UTR) and 3’ untranslated region (3’UTR) were amplified by PCR. The PCR products were purified and sequenced directly to detect Its gene mutation. Family members of DNA in the proband FII gene mutation region amplified after sequencing. Mutations were confirmed by restriction enzyme analysis. 103 healthy blood donors as controls. Results: Phenotype of proband was diagnosed as prothrombin deficiency (type Ⅰ). Three FII gene sequences were found in exon of FII, which were located in exon 2 Homozygous mutation A601G. Pedigree analysis shows that the proband’s father, mother and grandmother are A601G heterozygotes. CONCLUSIONS: Glu29 → GIy caused by homozygous missense mutation A601G is responsible for the hereditary prothrombin deficiency in this case. This is the first time in the world.