【摘要】 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。更多还原

【Abstract】 Objective To determine the quantity of the fetal mRNA in maternal peripheral blood and it’s variation in different gestation and gender.Methods From September to December of 2000,blood samples were collected and the fetal mRNA extracted from the pregnant women with the use of random primer in Public Health College,Harbin Medical University.Reverse transcription of mRNA into cDNA was carried out and cDNA was amplified by PCR with the special ε/γ primer used.“Gel Works System” were used to scan the electrophoresis image.Gel Works 1D can draft the quantitative curves to detect ε/γ gene band and fetal mRNA.Results RT PCR and agarose gel electrophoresis method,we detect ε/γ gene successfully in 50 samples,44 positive and 6 negative.The standard quantitative curve was:Y=-2 83×10 -5 X 2+0 06045X-2 265,R 2=0 9401.Mean of concentration of fetal mRNA in maternal peripheral blood was 20 79μg/L,maximum 59 72μg/L,and minimum 0 24μg/L.There was no statistically significant difference in different gestation and gender.Conclusion RT PCR method can directly detect fetal ε/γ gene from maternal peripheral blood.This can avoid the complicated method to separate fetal cells and can be easily accepted by pregnant women.Quantitative analysis of the fetal mRNA from maternal blood forms the base of studying the relationship between fetal mRNA contents and abnormal pregnancy.更多还原