论文部分内容阅读
目的探讨RASSF1A基因作为孕妇外周血中胎儿特异性标记物的可行性。方法随机选取43例的外周血样本,其中妊娠晚期孕妇标本40例、健康未妊娠女性标本3例,以及早期妊娠行流产的绒毛组织样本5例。利用甲基化敏感限制性内切酶PCR(MSRE-PCR)及荧光定量PCR的方法,检测血细胞、绒毛和血浆DNA的RASSF1A基因甲基化状态。结果健康未妊娠妇女血浆及孕妇血细胞中均未检出甲基化RASSF1A基因;绒毛组织中甲基化RASSF1A基因检出率为100%;孕妇血浆中甲基化RASSF1A基因检出率为92.5%。妊娠晚期孕妇血浆RASSF1A基因含量Ct平均值为30.43±2.37。结论在母体和胎儿DNA中,RASSF1A基因甲基化状态有明显差异,用实时荧光定量PCR可对孕妇血浆RASSF1A基因进行定量,提示了RASSF1A基因是无创产前诊断的一个潜在标记物。
Objective To investigate the feasibility of RASSF1A gene as a fetal specific marker in peripheral blood of pregnant women. Methods 43 cases of peripheral blood samples were randomly selected, including 40 cases of pregnant women in late pregnancy, 3 cases of healthy non-pregnant women, and 5 cases of abortion samples in early pregnancy. Methylation status of RASSF1A gene in blood, villi and plasma DNA was detected by methylation-sensitive restriction endonuclease PCR (MSRE-PCR) and real-time quantitative PCR. Results No methylated RASSF1A gene was detected in the plasma of pregnant women and pregnant women. The detection rate of methylated RASSF1A gene in villus was 100%. The detection rate of methylated RASSF1A gene in pregnant women was 92.5%. The average Ct of RASSF1A gene in pregnant women in the third trimester was 30.43 ± 2.37. Conclusion The methylation status of RASSF1A gene in maternal and fetal DNA is significantly different. Real-time quantitative PCR can be used to quantify the plasma RASSF1A gene in pregnant women, suggesting that RASSF1A gene is a potential marker in noninvasive prenatal diagnosis.