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为建立人周围血单核细胞载脂蛋白E基因表达检测方法 ,研究载脂蛋白E基因与儿童健康的关系 ,我们抽取 2 6例健康儿童外周静脉血 ,分离血单核细胞 ,抽提RNA ,采用逆转录—聚合酶链式反应检测载脂蛋白E基因表达 ,并以正常人cDNA作定量标准物 ,待测样品与定量标准物共扩增 ,计算出待测样本的个体的mRNA量。研究发现载脂蛋白E基因能在健康儿童周围血单核细胞表达 ,健康儿童载脂蛋白E基因表达量为 0 .37± 0 .15mol/molmRNA。表明采用竞争性逆转录—聚合酶链反应方法来检测人周围血单核细胞载脂蛋白E基因表达的分析方法快速、简便、灵敏、实用、可靠 ,且能准确定量 ,值得推广应用。
To establish a method to detect the expression of apolipoprotein E gene in peripheral blood mononuclear cells and study the relationship between Apolipoprotein E gene and children’s health, we extracted peripheral venous blood from 26 healthy children, isolated blood mononuclear cells, extracted RNA, The expression of apolipoprotein E gene was detected by reverse transcription polymerase chain reaction (RT-PCR). The normal human cDNA was used as a quantitative standard, and the sample and the quantitative standard were co-amplified to calculate the amount of mRNA of the individual to be tested. The study found that apolipoprotein E gene expression in peripheral blood mononuclear cells in healthy children, healthy children, the expression of apolipoprotein E gene was 0.37 ± 0. 15mol / molmRNA. The method of competitive reverse transcription polymerase chain reaction (RT-PCR) to detect apolipoprotein E gene expression in human peripheral blood mononuclear cells is rapid, simple, sensitive, practical and reliable, and can be accurately quantified.