摘要
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 。
To establish a quantitative assay for telomerase activity and analyze the telomerase activity in peripheral blood mononuclear cells(PBMNC) from patients with acute leukemia. A fluorescent dye, PicoGreen, was added to the products after telomere repeat amplification protocol. The samples were excited at 480 nm and the fluorescence emission intensity was measured at 520 nm using a spectrofluorometer. Telomerase activity was detected in PBMNCs from 20 cases of normal individuals and 25 patients with acute leukemia. The results showed that the fluorescence of PicoGreen binding to double stranded DNA specifically was enhanced with increase of DNA quantities. In conclusion, the method is rapid, simple and quantitative, the telomerase activities of PBMNCs from acute leukemia patients are significantly higher than that of the normal controls.
出处
《中国实验血液学杂志》
CAS
CSCD
2002年第3期191-194,共4页
Journal of Experimental Hematology
