LD-PCR检测FⅧ基因XbaⅠ位点多态性及其在血友病A携带者诊断中的应用  

Detection the polymorphism of Xba Ⅰ site in FⅧ gene using LD-PCR and its application on carrier detection for Hemophilia A

作　　者：刁戈[1] 马莉[1] 林方昭[1] 孙盼[1] 李长清[1] 肖小璞[1] 

机构地区：[1]中国医学科学院北京协和医学院输血研究所,四川成都610052

出　　处：《中国输血杂志》2013年第1期33-36,共4页Chinese Journal of Blood Transfusion

基　　金：2010年四川省科技支撑计划项目"血友病携带者及其产前基因诊断技术的研究"(2010SZ0130);2011年成都市科技计划项目"获得性血友病相关致病基因及分子诊断技术研究"(11PPYB037SF-289)

摘　　要：目的建立高特异性的针对凝血因子Ⅷ(FⅧ)基因内含子22中XbaⅠ多态性位点的连锁分析法,以用于血友病A家系携带者的诊断。方法采用长距离PCR(LD-PCR)方法,特异性地扩增FⅧ基因内含子22中的XbaⅠ位点所在区域,用限制性内切酶XbaⅠ酶切,对5个血友病A家系作家系连锁分析,并对其中可提供多态性信息的家系作携带者诊断。结果家系1中确诊女性携带者1名及1名正常女性成员;家系2中确诊1名女性携带者;而家系3、家系4和家系5该位点无法提供多态性信息。结论基于LD-PCR的FⅧ基因XbaⅠ位点连锁分析方法具有较高的准确性和可靠性,这对临床上诊断血友病A携带者具有积极的意义。Objective To conduct carrier detection in Hemophilia A families by establishing the linkage analysis meth- od specific for XbaⅠ polymorphic site in intron 22 of FⅧ gene. Methods The LD-PCR was applied to amplify the region which included the XbaⅠ site in intron 22 of FⅧ gene followed by XbaⅠ digestion. 5 Hemophilia A families were analyzed by linkage analysis. Carrier detection was performed if the XbaⅠ polymorphic site was informative. Results One female member was diagnosed as carrier and another one was normal in family 1. As to family 2 ,one female member was diagnosed as carrier. The XbaⅠ polymorphic sites were not informative in the other three families. Conclusion This improved linkage a- nalysis method for XbaⅠ site of FⅧ gene was more accurate and reliable, and it will be more meaningful for cartier detection of Hemophilia A in clinic.

关 键 词：血友病A FⅧ基因 LD—PCR 连锁分析 携带者诊断 

分 类 号：R446.1[医药卫生—诊断学] B554.11[医药卫生—临床医学]