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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: To compare the changes of PLAC4-DNA concentration in peripheral blood of normal pregnant women and pregnant women with 21-trisomy by a modified absolute quantitative PCR method.Methods The standard curve was constructed by using the long fragment amplicon instead of the plasmid, Standard curve equation was used to calculate the concentration of free PLAC4-DNA in the peripheral blood of two groups of pregnant women. Two independent samples t-test were used to analyze the differences between the two groups. Results The standard curve equation of the constructed amplicon was Y = -3.334X + 43.438 , R2 = 0.996, Eff% = 99.478%. According to the standard curve equation, the average concentration of PLAC4 in 300 normal pregnant women was 58.4 copies / μl, the average concentration of 20 pregnant women with trisomy 21 was 62.6 copies / μl, P = 0.439 by two independent samples t test. The improved long fragment amplicon construction standard curve has the advantages of simple operation, low cost and high accuracy, and provides an economical and convenient and effective reference method for absolute quantification. There was no significant difference in total free PLAC4-DNA in peripheral blood between normal pregnant women and gestational trisomy 21 pregnant women by absolute quantitative method.