胎儿脐静脉血凝血因子Ⅷ活性检测联合基因测序在甲型血友病产前诊断中的应用  

Combination of fetal cord blood coagulation factor Ⅷ activity assessment with gene sequencing in prenatal diagnosis of hemophilia A

作　　者：张燕[1] 王谢桐[1] 李红燕[1] 张雪芹[2] 

机构地区：[1]山东大学附属省立医院妇产科,济南250021 [2]山东省血液中心

出　　处：《中华围产医学杂志》2010年第5期394-397,共4页Chinese Journal of Perinatal Medicine

基　　金：基金项目：山东省科技发展计划项目（2004GG3202025）

摘　　要：目的探讨应用胎儿脐静脉血凝血因子Ⅷ活性（coagulation factor Ⅷ activity，FⅧ：C）检测结合基因诊断对甲型血友病高危胎儿进行产前诊断的临床可行性。方法对1996年10月至2008年10月的79例妊娠20－35周的甲型血友病高危且为男胎的孕妇，经腹B超引导下行胎儿脐静脉穿刺取血术。应用一期法和酶联免疫吸附法分别测定FⅧ：C及vWF抗原（von Willebrand factor antigen，vWF：Ag），对近1年来FⅧ：C〈10％的胎儿血（6例）采用长片段聚合酶链反应（long distance polymerase chain reaction，LD-PCR）检测内含子22倒位，对倒位阳性胎儿的孕母应用LD-PCR检测倒位携带者；对非倒位阳性胎儿及其孕母应用基因测序方法检测基因突变位点。结果79例孕妇血FⅧ：C为52％～139％，平均（99．60±28．10）％。胎儿血FⅧ：C〈10％者23例，平均（2．64±1．92）％，均引产；其中8例胎儿血FⅧ：C〈1％，15例胎儿血FⅧ：C〉1％。胎儿血FⅧ：C10％～30％者12例，平均（19．78±6．71）％，5例引产，7例继续妊娠。44例胎儿血FⅧ：C〉30％，平均（58．60±12．12）％，继续妊娠，所有51例未引产者生后随访至今尚未发现子代异常。在行基因诊断的6例（胎儿血FⅧ：C〈10％）中，内含子22倒位2例，另4例胎儿血基因测序尚未找到突变位点。结论应用胎儿血FⅧ：C及vWF：Ag浓度测定可以对甲型血友病高危孕妇进行产前诊断，结合LD-PCR技术可准确的对甲型血友病内含子22倒位者进行产前诊断及携带者诊断，但基因测序检测突变位点仍需要进一步研究。Objective To evaluate the assessment of fetal cord blood coagulation factor vm activity （Ⅷ ： c） combined with gene sequencing in prenatal diagnosis of high-risk fetuses of hemophilia A（HA）. Methods Percutaneous umbilical blood sampling was performed in 79 pregnant woman with high risk of HA and male fetuses at 20-35 weeks of gestation and maternal peripheral blood was also taken. The plasma FⅧ. C was detected by one-stage method, and the concentration of von Willebrand factor antigen （vWF： Ag） was determined by ELISA. Long distance polymerase chain reaction（LD-PCR） was used in 6 fetuses to detect the intron 22 inversion in F8 gene, and LD-PCR was also applied for mothers with fetal carriers of intron 22 inversion and gene sequencing for those mother with fetuses without intron 22 inversion. Results The maternal concentration of FⅧ： C of these 79 women was 52%-139% with an average of （99.60±28.10）%. For the fetuses, 23 were below 10% with an average of （2.64±1.92） %, eight of which were 〈1% and 15 between 1% and 10%. Among the 12 fetuses with the concentration of FⅧ ： C at 10 %-30%, averagely （ 19.78 ± 6.71） %, five were induced and the rest seven pregnancies and the other 44 pregnancies with fetal FⅧ：C over 30%, average （58.60±12. 12）%, were continued. All of the above 51 fetuses were followed up after birth without any abnormalities till present. Genetic diagnosis was offered for 6 fetuses, whose F Ⅷ ： C concentration 〈10 % （three 〉 1% and the other three 〈 1 % ）, and intron 22 inversion was detected in two and no mutation was found among the rest four. Conclusions Fetal blood FⅧ： C and vWF.. Ag concentrations determination may offer prenatal diagnosis of HA, and is accurate in detecting HA intron 22 inversion and its carriers when combined with LD-PCR.

关 键 词：血友病A 产前诊断 因子Ⅷ 胎血 

分 类 号：R714[医药卫生—妇产科学]