Fu Qi-hua, Wang Hong-li, Wang Ming-shan, Ding Qiu-lan, Wu Wen-man, Hu Yi-qun, Wang Xue-feng, Wang Zhen-yi
Shanghai Institute of Hematology, Ruijin Hospital, Shanghai Second Medical University, Shanghai 200025, China.
Zhonghua Yi Xue Za Zhi. 2003 Feb 25;83(4):312-5.
To discover the gene mutations of a pedigree with inherited factor V (FV) deficiency.
The activated partial thromboplastin time (APTT), prothrombin time (PT), FV activity (FV:C) and FV antigen test were adopted for phenotype diagnosis. The genomic DNA was extracted from the peripheral blood of the 16-year-old propositus, female. All the 25 exons and their flanks in the FV gene of the propositus were amplified by polymerase chain reaction (PCR). The PCR products were screened by direct sequencing and the mutations were further confirmed by restricted enzyme digestion. Six persons in the pedigree (grandfather, grandmother, father, mother, uncle, and aunt) were examined too. 108 healthy blood donors were used as controls.
The APTT, PT, FV:C, and FV:Ag of the propositus were 126.6s, 42.8s, 0.3% and 1.3% respectively. The Fbg and FII, FVII, FVIII, FIX, FX activities were in normal range. FV:C of the members of the pedigree was 36% - 70%, and the FV:Ag of the pedigree members was 26.4% - 45.3% that of the mixture of 30 normal plasma samples. Taking the GeneBank Z99572 sequence as the reference, totally five variations in the FV gene were found in the propositus. The mutations, A1348G and 4887 approximately 8delG, were traced to her father and her mother respectively. No 1348G-->T mutation was found in the 108 controls.
The FV deficiency of the propositus is caused by missense mutation of G1348T and frameshift mutation of 4887 approximately 8delG, which haven't been identified previously.
发现一个遗传性因子V（FV）缺乏家系的基因突变。
采用活化部分凝血活酶时间（APTT）、凝血酶原时间（PT）、FV活性（FV:C）及FV抗原检测进行表型诊断。从16岁先证者（女性）外周血中提取基因组DNA。采用聚合酶链反应（PCR）扩增先证者FV基因的全部25个外显子及其侧翼序列。PCR产物经直接测序筛选，并用限制性酶切进一步确认突变。对该家系中的6人（祖父、祖母、父亲、母亲、叔叔和阿姨）也进行了检测。1０８名健康献血者作为对照。
先证者的APTT、PT、FV:C和FV:Ag分别为126.6秒、42.8秒、0.3%和1.3%。纤维蛋白原及FII、FVII、FVIII、FIX、FX活性均在正常范围。家系成员的FV:C为36% - 70%，家系成员的FV:Ag为30份正常血浆混合样本的26.4% - 45.3%。以GeneBank Z99572序列为参照，在先证者FV基因中共发现5个变异。其中A1348G和4887约8delG突变分别来自其父亲和母亲。108名对照中未发现1348G→T突变。
先证者的FV缺乏是由G1348T错义突变和4887约8delG移码突变引起的，这两种突变此前尚未见报道。
