Hou Li-hong, Xie Fei, Liu Xiu-e, Zhang Li, Guo Yan-li, Dong Chun-xia, Li Zhi-ting, Yang Bo, Yang Lin-hua
The Second Hospital of Shanxi Medical University, Taiyuan 030001, China.
Zhonghua Xue Ye Xue Za Zhi. 2003 Sep;24(9):455-9.
To investigate the gene defect in a hereditary coagulation factor V (FV) deficiency family.
The plasma FV actigen was measured by one-stage clotting assay. The FV antigen was assayed by Biotin-Avidin enzyme linked immunosorbent assay (BA-ELISA). The full length of exon 1 to exon 25 and the 5' untranslated sequence of FV genomic DNA were analyzed by polymerase chain reaction (PCR) and direct sequencing of the amplified fragments, meanwhile the defect was identified by T/A cloning sequencing.
The plasma coagulant activity and amount of FV of the proband were marked deficient (1% and 1.54%, respectively). DNA sequence analysis for the proband revealed a causative mutation in a heterozygous status. It was one base pair deletion in exon 4 at nucleotide 675 inherited from her mother.
A novel mutation in the FV gene was identified in the proband with congenital FV deficiency. The mutation was 675delA in exon 4 resulting in a frameshift and a premature termination codon.
研究一个遗传性凝血因子V（FV）缺乏家族的基因缺陷。
采用一期凝血法测定血浆FV活性抗原。用生物素-抗生物素蛋白酶联免疫吸附测定法（BA-ELISA）检测FV抗原。通过聚合酶链反应（PCR）和对扩增片段的直接测序分析FV基因组DNA外显子1至外显子25的全长及5'非翻译序列，同时通过T/A克隆测序鉴定缺陷。
先证者的血浆凝血活性和FV量显著缺乏（分别为1%和1.54%）。对先证者的DNA序列分析显示存在杂合状态的致病突变。这是外显子4中第675位核苷酸处的一个碱基对缺失，遗传自她的母亲。
在先证者先天性FV缺乏中鉴定出FV基因的一个新突变。该突变是外显子4中的675delA，导致移码和提前终止密码子。
