摘要
目的 探讨先天性凝血因子Ⅴ (FⅤ )缺乏症的分子发病机制。方法 利用发色底物法检测血浆FⅤ凝血活性 ,采用PCR产物直接测序或T A克隆后测序、限制性酶切分析先证者FⅤ基因 ,并进行了一个家系和FⅤ基因多态性频率研究。结果 先证者 8号内含子 3′端剪接位点存在AG→GG的突变 ,这在家系和 10 0例正常对照人群的研究中得到证实。结论 FⅤ基因 8号内含子 3′端剪接位点的突变与先天性凝血因子Ⅴ缺乏症的发病有关。
Objective To explore the molecular mechanism involved in patient with congenital FⅤ deficiency. Methods Activity of FV was determined by biochemical method. The PCR products of FⅤ gene was analyzed by DNA sequencing directly or cloned into T vector prior to DNA analysis. The mutation of FⅤ gene in proband and his family numbers was analysed by restriction enzyme analysis. Its occurrence was investigated in the control group. DNA was extracted from the peripheral blood mono1nuclear cells of the proband, male, 18 years old, and his parents. The PCR products were analyzed by direct sequencing or cloned into T vector prior to DNA analysis. One hundred patients with different kind of hemotopathy were used as controls. Results A single point mutation, AG→GG was found at position 3′ splice site of intron 8 of the proband. This mutation was confirmed by family screening. Conclusion A single point mutation, AG→GG at position 3′ splice site of intron 8 mutation of FⅤ gene is related to the pathogenesis of congenital FV deficiency.
出处
《中华医学杂志》
CAS
CSCD
北大核心
2003年第1期24-26,共3页
National Medical Journal of China
