长距离PCR技术检测重型血友病甲Ⅷ因子基因倒位的进一步研究及推广应用  

Further Study on Long Distance PCR Detection of Factor Ⅷ Inversion Causing Severe Hemophilia A and Its Applications

作　　者：梁燕[1] 刘敬忠[1] 肖白[1] 季林祥[2] 张跃进[3] 赵翠兰[4] 王勇[5] 林雯[6] 

机构地区：[1]首都医科大学附属北京红十字朝阳医院北京基因诊断实验室 [2]中国医学科学院血液病研究所 [3]东南大学职工医院 [4]北京市宣武区椿树医院北京市京安疑难病研究所 [5]北京市宣武区椿树医院北京市京安疑难病研究所 [6]同济医科大学附属协和医院

出　　处：《高技术通讯》1999年第9期42-45,共4页Chinese High Technology Letters

摘　　要：将已研究成功的长距离ＰＣＲ（ＬＤＰＣＲ） 技术［３］在热循环条件、模板ＤＮＡ用量及ｄＮＴＰ用量等方面进行了进一步优化和探索。应用优化后的ＬＤＰＣＲ技术检测了来自天津、江苏、北京、湖北、四川等省市１０８个重型血友病甲家系共１５７ 份ＤＮＡ（患者１０５人， 家属５２人）， 发现４７例患者（或家系） 有凝血因子Ⅷ（ＦⅧ） 基因倒位， 占重型患者的４４％ ； 查出基因倒位携带者３０ 余名。用ＬＤＰＣＲ检测ＦⅧ基因倒位的技术可以准确、简便、快速地直接进行重型血友病甲的基因诊断、携带者检测及产前诊断。To detect Factor Ⅷ (FⅧ)gene inversion leading to severe Hemophilia A and carrier, a new technique based on the Long Distance Polymerase Chain Reaction (LD PCR) was developed and improved further. One hundred and eight of severe HA patients or families were detected by using the LD PCR, and the results show that: 47 were the FⅧ gene inversion; more than 30 cases of carriers of the Inversion were diagnosed also, which were 44% of total number of the severe HA. Compare with the Southern Blot method, the LD PCR shows more advantages, such as higher sensitivity, less DNA required, simpler and faster, and no radiactive isolope was required.

关 键 词：LD-PCR 血友病甲 Ⅷ因子 基因倒位 

分 类 号：R554.1[医药卫生—血液循环系统疾病]