【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LD-PCR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Bc1ⅠPCR/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0.9%~90%(平均41.7%±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;Bc1ⅠPCR/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LD-PCR和Bc lⅠPCR/RFLP基因分析技术可提高HA携带者的诊断率。更多还原

【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 one-stage.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 0.9%～90%,60%～77% and 58%～74%,respectively.The average were 41.7%±25%,69%±16% and 66%±13%,respectively.19 cases of 24 fetuses FVⅢ:C were higher than 34%,and 5 fetuses were below 5%.LD-PRC 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 LD-PCR and Bcl Ⅰ PCR/RFLP can improve the detection of hemophilia A carrier.更多还原