论文部分内容阅读
目的 建立一种重型血友病 A(hemophilia A,HA)携带者检测和产前诊断方法。方法 选择有重型 HA家族史的 2 5例 HA携带者 ,采用长距离 DNA扩增 (long distance- PCR,L D- PCR)技术 ,直接检测是否存在凝血因子 (factor ,F )基因倒位。对 L D- PCR基因诊断阳性携带者于妊娠 2 0～ 2 4周抽取夫妻静脉血和胎儿脐静脉血 ,应用 Bigg’s一期法检测血浆凝血因子 F 活性 (F ∶C) ,EL ISA法检测血浆血管性血友病因子 (von Willbrand factor,v WF)浓度 ,进一步应用 L D- PCR技术进行产前诊断。应用DNA测序技术验证 L D- PCR结果。结果 在 2 5个无亲缘关系的重型 HA家系中查出 F 基因倒位携带者 8例 ,其中对 4例 L D- PCR基因诊断阳性的携带者进行了产前诊断 ,2例确诊胎儿为 HA患者而建议终止妊娠 ,另 2名为正常胎儿 ,随访 1年 ,小儿发育正常。结论 应用 L D- PCR技术结合携带者外周静脉血及胎儿脐血血浆 F ∶ C和 v WF∶Ag浓度可准确、快速进行重型 HA患者的诊断及其携带者的产前诊断 ,防止血友病患儿的出生
Objective To establish a method for the detection and prenatal diagnosis of hemophilia A (HA) carriers. Methods Twenty-five cases of HA carriers with a family history of severe HA were selected to detect the presence or absence of the in-position factor F gene using long distance-PCR (L-PCR). Positive pregnant women diagnosed by L D-PCR gene were randomly divided into two groups: pregnant women and their umbilical blood samples were collected from 20 to 24 weeks of gestation. Bigg’s first phase assay was used to detect plasma F factor F (F: C) Von Willebrand factor (vWF) concentration, and further use of L-PCR technology for prenatal diagnosis. The DNA sequencing technique was used to validate L-PCR results. Results Eight out of twenty-five unrelated heavy HA pedigrees were found to have an inverted F-carrier. Among them, four were diagnosed as prenatal carriers of L-PCR gene diagnosis and two were diagnosed as HA The patient and the proposed termination of pregnancy, the other two normal fetuses, followed up for 1 year, children with normal development. Conclusion The combination of L-PCR with peripheral venous blood of carriers and fetal plasma F: C and WF: Ag concentrations can accurately and rapidly diagnose and carry out the prenatal diagnosis of patients with severe HA and prevent hemophilia Children’s birth