论文部分内容阅读
目的本文旨在探索使用荧光定量PCR方法对孕妇全血和血浆中DNA含量进行动态分析,从而为利用孕母血浆循环DNA进行无创产前诊断奠定基础。方法提取60例早、中、晚孕期孕妇全血基因组DNA和血浆循环DNA,应用实时荧光定量聚合酶链式反应(FQ-PCR)技术测定不同孕期的孕妇全血和血浆中保守基因(β-actin)的含量,比较不同孕期孕妇全血和血浆DNA含量的变化。结果 (1)早、中、晚孕期孕妇全血DNA含量Ct平均值为18.44±0.56828、18.35±0.93394、18.15±0.57410,3组比较没有显著性差异(P>0.05);(2)孕妇早、中、晚期孕组血浆DNA含量Ct平均值25.59±1.17673、25.50±1.59892、24.35±2.24301,早、中期孕组血浆DNA没有显著性差异(P>0.05),早、晚期和中、晚期孕组血浆DNA在两者之间有显著性差异(P<0.05);(3)排除全血DNA干扰,早、中、晚孕期孕妇血浆中DNA相对定量值为0.007401、0.007401、0.013602。结论用实时荧光定量PCR的方法可以检测到孕妇血浆中循环DNA的含量变化,随孕周的增加而逐渐增加,可以利用其进行无创产前诊断。
Objective This study aimed to explore the use of fluorescence quantitative PCR method for pregnant women, whole blood and plasma DNA content in dynamic analysis, so as to lay the foundation for the use of maternal plasma DNA for noninvasive prenatal diagnosis. Methods Whole blood genomic DNA and plasma DNA were extracted from 60 pregnant women in early, middle and late pregnancy. Real-time quantitative polymerase chain reaction (FQ-PCR) was used to detect the levels of conserved genes in whole blood and plasma (β- actin) content in different pregnant women during pregnancy compared whole blood and plasma DNA content changes. Results (1) The average Ct of DNA in pregnant women in early, middle and late pregnancy was 18.44 ± 0.56828, 18.35 ± 0.93394 and 18.15 ± 0.57410, respectively. There was no significant difference between the three groups (P> 0.05). (2) In middle and late pregnancy group, the plasma Ct levels in plasma were 25.59 ± 1.17673,25.50 ± 1.59892,24.35 ± 2.24301, there was no significant difference in plasma DNA between early pregnancy and intermediate pregnancy (P> 0.05) (P <0.05). (3) Excluding whole blood DNA interference, the relative quantitative value of DNA in pregnant women in early, middle and late pregnancy was 0.007401, 0.007401 and 0.013602, respectively. Conclusion The real-time PCR method can detect the change of circulating DNA in pregnant women’s plasma and increase gradually with the increase of gestational age, so it can be used for non-invasive prenatal diagnosis.