用改良绝对定量法检测正常孕妇与妊娠21三体胎儿孕妇外周血游离核酸浓度  

An improved absolute quantification PCR( AQ—PCR) method for analyzing the total cell-free DNA in the serum of pregnant women carrying euploidy and trisomy 21 fetuses

作　　者：黄睿[1] 王树玉[2] 马延敏[2] 贾婵维[2] 

机构地区：[1]首都医科大学附属复兴医院 [2]首都医科大学附属北京妇产医院,100026

出　　处：《中国优生与遗传杂志》2014年第1期29-31,共3页Chinese Journal of Birth Health & Heredity

基　　金：首都医学发展科研基金资助(2009-3130)

摘　　要：目的运用一种改良的绝对定量PCR法比较分析正常孕妇组与妊娠21-三体孕妇组外周血中游离PLAC4-DNA浓度的变化;方法采用长片段扩增子代替质粒构建标准曲线,根据生成的标准曲线方程计算出两组孕妇外周血中游离PLAC4-DNA的浓度值,经两独立样本t检验分析两组结果差异;结果长片段扩增子构建的标准曲线方程:Y=-3.334X+43.438,R2=0.996,Eff%=99.478%。根据标准曲线方程得出,300例正常孕妇组PLAC4平均浓度:58.4 copies/μl,20例妊娠21三体孕妇组平均浓度:62.6 copies/μl,,经两独立样本t检验,P=0.439;结论改良的长片段扩增子构建标准曲线具有操作简便,成本低,准确性高的优点,可为绝对定量提供一种经济简便且有效的参考方法。通过绝对定量法检测得出正常组孕妇与妊娠21-三体孕妇组外周血中总的游离PLAC4-DNA的变化无统计学差异。Objective： Analysis the change of cell free PLACA - gene DNA concentration in serum of pregnant women carrying eu- ploidy and trisomy 21 fetuses by using a new kind of improved absolute quantification PCR （ AQ - PCR） method; Methods ： Using long fragment amplicon instead of plasmid vector to build standard curve for AQ - PCR, according to which generate the standard curve e- quation to calculate the free DNA density of two groups, analyzing the results of two groups by the two independent samples t - test; Results ： Standard curve equation by long fragment amplicon method is as follow： Y = - 3. 334X ＋ 43.438, R^2 = 0. 996, Eft% = 99. 478%. According to the standard curve equation, the average cell free fetal DNA （ff-DNA） concentration is 58.4 copies/μl in 300 cases of normal pregnancy women, and is 62. 6 copies/μl in 20 eases of trisomy 21 pregnancy women, which are no significant differences （ P = 0. 439 〉 0. 05 ） ; Conclusion ： The improved long fragment amplieon method with the features of easy operation, low cost, and high accuracy, can provide an more effective referenced method for AQ - PCR. And it is not applicable for non - invasive pregnancy diagnosis by detecting the change of cell free PLACA - gene DNA levels in serum of pregnant women carrying euploidy and trisomy 21 fetuses through AQ - PCR method.

关 键 词：绝对定量PCR 游离PLAC4-DNA 21三体 

分 类 号：R714.55[医药卫生—妇产科学]