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目的 分析孕妇外周血中胎儿mRNA的含量及不同孕期不同性别胎儿mRNA含量的变化。方法 2 0 0 0年 9～ 12月哈尔滨医科大学公共卫生学院等单位提取孕妇全血mRNA ,经体外逆转录后 ,应用ε/γ引物引导扩增胎儿特异血红蛋白基因片段 ,同时用不同剂量标准mRNA经逆转录及PCR扩增后 ,琼脂糖凝胶电泳图用凝胶成像系统扫描成像 ,用GelWorks 1D软件分析凝胶图像 ,绘制定量标准曲线 ,对样品进行定量分析。结果 5 0例样品中 4 4例扩增出了胎儿ε/γ基因片段 (2 74bp) ,6例为阴性。定量标准曲线公式 :Y =- 2 83× 10 -5X2 +0 0 6 0 4 5X - 2 2 6 5 ,R2 =0 94 0 1,胎儿mRNA在孕妇外周血中的质量浓度均值为 2 0 79μg/L ,最大值 5 9 72 μg/L ,最小值 0 2 4 μg/L。不同孕期及不同性别胎儿mRNA含量统计学上差异无显著性意义。 结论 应用反转录聚合酶链反应 (RT PCR)方法可直接自孕妇外周血中扩增出胎儿ε/γ球蛋白基因片断 ,对孕妇全血中的胎儿mRNA进行定量分析 ,为探索胎儿mRNA含量变化与异常妊娠的关系奠定了基础。
Objective To analyze the content of fetal mRNA in peripheral blood of pregnant women and the changes of mRNA levels in different sexes of fetus during pregnancy. Methods From September to December 2001, Harbin Medical University School of Public Health and other units to extract whole blood mRNA of pregnant women, after in vitro reverse transcription, ε / γ primers were used to guide the amplification of fetal hemoglobin gene fragments, with different doses of standard mRNA After reverse transcription and PCR amplification, agarose gel electrophoresis images were scanned with a gel imaging system. GelWorks 1D software was used to analyze the gel images and quantitative standard curves were drawn to quantitatively analyze the samples. Results Fetal ε / γ gene fragment (2 74bp) was amplified in 44 cases of 50 samples, and 6 cases were negative. Quantitative standard curve formula: Y = - 2 83 × 10 -5X2 +0 0 6 0 4 5X - 2 2 6 5, R2 = 0 94 0 1, the average concentration of fetal mRNA in peripheral blood of pregnant women is 2 0 79 μg / L, the maximum value of 5 9 72 μg / L, the minimum value of 0 2 4 μg / L. There was no significant difference in the mRNA levels of fetus in different pregnancy and gender. Conclusion RT-PCR can be used to amplify the fetus ε / γ globin gene fragment directly from the peripheral blood of pregnant women, and quantitatively analyze the fetal mRNA in whole blood of pregnant women. In order to explore the fetal mRNA content Changes and abnormal pregnancy laid the foundation for the relationship.