Wang Wenjing, Huang Shan, Li Jingjing, Rui Kai, Zhang Jian-Rong, Zhu Jun-Jie
State Key Laboratory of Analytical Chemistry for Life Science and Collaborative Innovation Center of Chemistry for Life Sciences, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210093, China.
School of Pharmacy, Ningxia Medical University, Yinchuan 750004, China.
Sci Rep. 2016 Mar 24;6:23504. doi: 10.1038/srep23504.
The strong correlation between cancer and telomerase activity has inspired the development of new strategies to evaluate telomerase activity. Here, a personal glucose meter (PGM) system that uses DNA-based machine amplification to detect telomerase in cancer cells is reported. In this assay, telomerase elongation products are amplified in the form of another type of product by a DNA-based machine. This process can only be activated by the hybridization of the extended telomerase substrate (TS) probe and the complementary primer in the presence of telomerase. The obtained products are then transformed to glucose-related signals via a three-component assay, which enables the simple use of a PGM to indirectly quantify the telomerase activity. The proposed method realizes sensitive telomerase activity detection down to 20 HeLa cells with a significantly enhanced dynamic range. Additionally, short telomerase elongation products, such as telomerase substrate probes with two repetitive sequences, that cannot be detected using the most widely used telomeric repeat amplification protocol assay were detected.
癌症与端粒酶活性之间的强相关性激发了评估端粒酶活性新策略的发展。在此，报道了一种使用基于DNA的机器扩增来检测癌细胞中端粒酶的个人血糖仪（PGM）系统。在该检测中，端粒酶延伸产物通过基于DNA的机器以另一种产物的形式被扩增。此过程仅在端粒酶存在的情况下，通过延伸的端粒酶底物（TS）探针与互补引物的杂交才能被激活。然后，通过一个三组分检测将获得的产物转化为与葡萄糖相关的信号，这使得能够简单地使用PGM间接定量端粒酶活性。所提出的方法实现了对低至20个HeLa细胞的敏感端粒酶活性检测，且动态范围显著增强。此外，还检测到了短的端粒酶延伸产物，例如具有两个重复序列的端粒酶底物探针，而这些产物使用最广泛的端粒重复序列扩增协议检测无法检测到。
