论文部分内容阅读
本文介绍一种端粒酶检测方法，该方法是将 Ｔ Ｒ Ａ Ｐ法在 Ｐ Ｃ Ｒ 过程中引入３ Ｈｄ Ｃ Ｔ Ｐ，结合液闪计数ｃｐｍ 半定量分析端粒酶活性。应用该方法检测端粒酶阳性的 Ｃ Ｎ Ｅ２ 细胞和部分组织标本及 Ｒ Ｎａｓｅ Ａ 或加热预处理后的对照标本，并与 Ｔ Ｒ Ａ Ｐ 法相比较，结果显示 Ｃ Ｎ Ｅ２ 细胞端粒酶阳性，１０ 个 Ｃ Ｎ Ｅ２ 细胞中仍可检到端粒酶，组织样品的端粒酶与文献值基本一致；放射性活性计数（ｃｐｍ） 与 Ｃ Ｎ Ｅ２ 细胞抽提液中端粒酶活性具有良好的线性关系；用 Ｒ Ｎａｓｅ Ａ 或加热处理后标本为阴性，ｃｐｍ 值接近阴性对照；批内和批间变异度分别为１１ ．６８ ％ 和２０９９ ％ 。该方法不需使用聚丙烯酰胺凝胶电泳（ Ｐ Ａ Ｇ Ｅ） 和放射自显影，简便快速，当天可观察结果，并具有灵敏度高、特异性和批内重复性好之特点。
This article describes a method for detecting telomerase. The method is to introduce 3 H-d C T P during the PRR process using the T R A P method and semiquantitatively analyze the telomerase activity with the liquid scintillation counter cpm. This method was used to detect telomerase-positive CNE2 cells and some tissue specimens and R Nase A or heated pretreated control specimens. Compared with T R A P method, the results showed that CN N E2 cells were positive for telomerase. Telomerase was still detectable in 10 C N E2 cells. The telomerase activity of the tissue samples was almost the same as the literature value; the radioactivity activity count (cpm) was well linearized with the telomerase activity of CN E2 cell extracts. Relationships; specimens with R Nase A or heat treatment were negative, cpm values were close to negative controls; intra-assay and inter-assay variability were 11, respectively. 68% and 2099%. This method does not require the use of polyacrylamide gel electrophoresis (PGAE) and autoradiography. It is simple and rapid, and can be observed on the day. It has high sensitivity, specificity, and good intra-assay reproducibility.