摘要
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 。
Objective To establish an effective laboratory examination system for carrier detection and prenatal diagnosis of haemophilia A (HA). Methods Twenty-five carriers of severe HA were directly detected by long-distance PCR(LD-PCR) in search of the factor Ⅷ(FⅧ) gene inversion. Prenatal diagnosis was carried out using pregnant woman's venous blood sample,husband's venous blood sample and fetal navel venous sample at 20-24 weeks of gestation. The plasma coagulation factor Ⅷ activity (FⅧ:C) was detected by one-stage method. The concentration of von Willbrand factor (vWF) was assayed by ELISA. Prenatal diagnosis was finally made by LD-PCR. The results of LD-PCR were proved by DNA sequencing. Results Eight out of 25 cases were diagnosed as having FⅧ gene inversion. Four of these 8 carriers underwent the LD-PCR for prenatal diagnosis, and 2 of them had to terminate pregnancy because their fetuses were diagnosed as having HA. The other two carriers were finally diagnosed to have normal fetuses by combined use of LD-PCR with plasma FⅧ:C, vWF in pregnant woman's venous blood, husband's venous blood and fetal navel venous blood, and the one-year follow-up study demonstrated that the babies were normal and living well. Conclusion LD-PCR technique was adopted in this study to detect the factor Ⅷ gene inversion; it could accurately and rapidly diagnose the severe cases of HA and could be used for the HA carriers in need of pregnant diagnosis.
出处
《中华医学遗传学杂志》
CAS
CSCD
2004年第5期505-507,共3页
Chinese Journal of Medical Genetics
基金
山东省自然科学基金 (Y2 0 0 2 C0 2 )
山东省卫生厅青年基金 (2 0 0 1 CA2 DMA1 )~~
