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为建立一种简便、定量检测端粒酶活性的方法应用于白血病病人外周血单个核细胞 (MNC)端粒酶活性分析 ,在应用端粒重复序列扩增法后在扩增产物内加入荧光染料PicoGreen ,并在激发光 4 80nm和发射光 5 2 0nm下检测荧光强度。对 2 0例正常人和 2 5例急性白血病病人外周血单个核细胞的端粒酶活性进行了定量分析。结果表明 ,PicoGreen能特异结合双链DNA ,荧光强度随双链DNA量的增加而增加。结论 :该方法简便、快速 ,能定量 ,可用于急性白血病病人外周血单个核细胞端粒酶活性分析 ,并提示急性白血病病人外周血单个核细胞的端粒酶活性明显高于正常人
To establish a simple and quantitative method for detecting telomerase activity in leukemia patients with peripheral blood mononuclear cells (MNC) telomerase activity analysis, the telomerase repeat amplification method after the application of fluorescent dye PicoGreen, and in the excitation light 480nm and emission light 5220nm fluorescence intensity was detected. The telomerase activity of peripheral blood mononuclear cells in 20 normal subjects and 25 acute leukemia patients was quantitatively analyzed. The results showed that PicoGreen could specifically bind double-stranded DNA, and the fluorescence intensity increased with the amount of double-stranded DNA. Conclusion: The method is simple, rapid and accurate and can be used to analyze the telomerase activity of peripheral blood mononuclear cells in patients with acute leukemia and suggests that the telomerase activity of peripheral blood mononuclear cells in patients with acute leukemia is significantly higher than that in normal people