甲型血友病的基因诊断研究  

Study on genetic diagnosis of hemophilia A

作　　者：尹璐[1] 宋淑霞[1] 连伟光[1] 栗彦宁[1] 郑龙[1] 刘健敏[1] 尤红煜[1] 王俊霞[1] 

机构地区：[1]河北省实验动物重点实验室 河北医科大学 分子生物学研究室,河北石家庄050017

出　　处：《临床荟萃》2014年第4期378-380,F0003,共4页Clinical Focus

基　　金：河北省科技攻关计划项目(11276102D)

摘　　要：目的建立甲型血友病的基因诊断方法。方法对33例甲型血友病患者以一期法检测血浆凝血因子FⅧ(F8)活性。采用长距离聚合酶链式反应扩增法(LD-PCR)进行FⅧ内含子22倒位检测,并对反应体系进行优化;聚合酶链式反应(PCR)技术检测内含子1倒位,凝胶成像技术分析扩增产物。捕获测序技术检测是否存在其他突变类型。结果 33例患者的FⅧ活性检测结果均小于1%,为重型;LD-PCR技术检测出13例患者存在Int22倒位,倒位发生率为39.4%;Int1倒位患者1人,发生率3%;对13例内含子22倒位患者进行测序比对,未发现其他基因突变。结论 LD-PCR技术和多重PCR技术可以有效的检测重型甲型血友病FⅧ的基因倒位。Objective To estahlish a method for direct genetic diagnosis on hemophilia A（HA）. Methods The concentration of FⅧ （FⅧ ： C） was detected with one stage method in 33 HA patients. Detection of factor Ⅷ gene inversion among 33 patients with severe hemophilia A was performed by long distance polymerase chain reaction（PCR） and 0.6 % agarose gel electrophoresis. Multiple-PCR was used to detect intron 1 inversions respectively,seeking other mutations with sequencing technology. Results All 33 HA who were diagnosed by FⅧ .. C were severe. Intron 22 inversion was detected in 13 HA patients,accounting for 39.4% （13/33）. And intron 1 inversion was detected in 1 HA patient,for 3.0% （1/33）. Other mutation was not found by sequencing technology. Conclusion LD- PCR and multiple- PCR are effective method for detecting factor W gene inversion.

关 键 词：血友病A 因子Ⅷ 序列倒位 多重聚合酶链式反应 

分 类 号：R554.1[医药卫生—血液循环系统疾病]