脐带血用于血友病A产前诊断的研究_医学论文.docVIP

脐带血用于血友病A产前诊断的研究_医学论文 脐带血用于血友病A产前诊断的研究_医学论文 作者：张雪芹 陈会昌 李文超 滕彬 张萍 【关键词】 血友病A 　　【摘要】 目的 探讨应用脐带血对血友病A（HA）进行产前诊断的可行性及准确性。方法 24例HA携带者妊娠20～２４周时在超声引导下抽取脐血，用一期法检测孕妇外周血及胎儿脐血FVⅢ：Ｃ，用ELISA方法检测FVⅢ：ＣＡｇ及ｖＷＦ；应用LDＰＣＲ技术对携带者及其胎儿进行内含子22倒位检测，对非倒位者应用Bｃ１　Ⅰ PCR／ＲＦＬＰ分析技术进行间接基因诊断。结果 24例携带者血浆FVⅢ：Ｃ，ＦＶⅢ：ＣＡｇ及ｖＷＦ分别为36％～１５０％(平均79%±２２％)，90%～１３２％(平均102%±１８％），92％～１４１％(平均107%±１６％）；２４例胎儿血浆FVⅢ：Ｃ，ＦＶⅢ：ＣＡｇ及ｖＷＦ分别为09％～９０％(平均417%±２５％），６０％～７７％（平均69%±１６％），５８％～７４％（平均66%±１３％），19例胎儿FVⅢ：Ｃ高于34%，5例胎儿FVⅢ：Ｃ低于5%；内含子22倒位胎儿1例，倒位携带者3例；Bc1 Ⅰ　ＰＣＲ／ＲＦＬＰ连锁分析诊断中型HA胎儿2例。结论 脐静脉穿刺取血测定FVⅢ：Ｃ是产前诊断HA的可行性手段，应用LDＰＣＲ和Bcl Ⅰ　ＰＣＲ／ＲＦＬＰ基因分析技术可提高HA携带者的诊断率。 　　【关键词】 血友病A；产前诊断；脐带穿刺 　　 　　【Ａbstract】 Objective Ｔo investigate the feasibility and accuracy of hemophilia A prenatal diagnosis by fetal blood sampling.Ｍethods Ｃordocentesis were performed in 24 pregnant hemophilia A carriers during 20~２４　ｇｅｓｔａｔｉｏｎａｌ　ｗｅｅｋｓ．Ｔｈｅ　ｐｌａｓｍａ　ｃｏａｇｕｌａｔｉｏｎ　ｆａｃｔｏｒ　Ⅷ activity (FVⅢ：Ｃ） of carrier’s peripheral blood and fetus umbilical blood were detected by Bigg’s onestage.Ｍethod The concentration of von Willbrand Factor (vWF) and FVⅢ：CAｇ　were assayed by ELISA.Intron 22 inversion was directly examined by long distance polymerase chain reaction,indirectly gene diagnosis was done by utilizing heredity linkage analysis of factor Ⅷ　ｉｎｔｒａｇｅｎｉｃ　Ｒｅｓｔｒｉｃｔｉｏｎ　Ｆｒａｇｍｅｎｔ　Ｌｅｎｇｔｈ　Ｐｏｌｙｍｏｒｐｈｉｓｍ　（ＲＦＬＰ）　ｏｆ　Ｂｃｌ　Ⅰ．Ｒesults ＦＶⅢ：Ｃ，ＦＶⅢ：ＣＡｇ　ａｎｄ　ｖＷＦ　ｏｆ　２４　ｃａｒｒｉｅｒｓ　ｗｅｒｅ　　３６％～１５０％，９０％～132%　ａｎｄ　９２％～１４１％，respectively.The average were 79%±２２％，102%±１８％ 　ａｎｄ　１０７％±１６％，ｒｅｓｐｅｃｔｉｖｅｌｙ．ＦＶⅢ：Ｃ，ＦＶⅢ：ＣＡｇ　ａｎｄ　ｖＷＦ　ｏｆ　２４　ｆｅｔｕｅｓｅ　ｕｍｂｉｌｉｃａｌ　ｂｌｏｏｄ　ｗｅｒｅ　０９％～９0%，60%～７７％　ａｎｄ　５８％～７４％，respectively.The average were 41７％±２５％，６９％±１６％　ａｎｄ　６６％±１３％，respectively.19 cases of 24 fetuses FVⅢ：Ｃ　were higher than 34%,and 5 fetuses were below 5%.LDＰＲＣ showed that 1 fetus and 3 carriers were FVⅢ ⅣＳ２２，　ａｎｄ　２　ｆｅｔｕｓｅｓ　ｗｅｒｅ　ｄｉａｇｎｏｓｅｄ　ＨＡ　ｂｙ　Ｂｃｌ　Ⅰ　ＰＣＲ／ＲＦＬＰ．Ｃonclusion Ｉt is an effective method in prenatal diagnosis of hemophilia A to detect FVⅢ：Ｃ of fetuses umbilical blood.The technology of LDＰＣＲ and Bcl Ⅰ PCR/RFLP can improve the detection of