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 210023, China.
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 210023, China.
Talanta. 2018 Feb 1;178:594-599. doi: 10.1016/j.talanta.2017.09.070. Epub 2017 Sep 28.
A simple yet powerful polymerase chain reaction (PCR)-free strategy for visualized assay of human telomerase activity was reported in this work. Gold nanoparticles (AuNPs) based colorimetric strategy was applied with well-designed enzyme-aided cyclic amplification. Briefly, the detection relies on the elongated primers of telomerase substrate (TS) induced by telomerase, which open the hairpin DNA and hybridize with linker DNA, the trigger of AuNPs aggregation. Nicking endonuclease was added in the sensing system, which cleaved linker DNA after hybridization and released complimentary strand for cyclic hybridization with linker DNA, resulted in high sensitivity for the detection of telomerase. Down to 25 HeLa cells with high expression of telomerase could be recognized. The proposed strategy provides a good platform for the determination of telomerase activity, differentiation of cancer cell lines from normal cell line and screening of telomerase-targeted anticancer drugs.
本工作报道了一种简单而强大的聚合酶链反应（PCR）免费策略，用于可视化测定人端粒酶活性。该策略应用了基于金纳米粒子（AuNPs）的比色策略，并结合了精心设计的酶辅助循环扩增。简而言之，该检测依赖于端粒酶诱导的端粒酶底物（TS）的延伸引物，该引物打开发夹 DNA 并与连接 DNA 杂交，连接 DNA 是 AuNPs 聚集的触发物。在传感系统中加入了切口内切酶，它在杂交后切割连接 DNA，并释放互补链与连接 DNA 进行循环杂交，从而实现了对端粒酶的高灵敏度检测。可以检测到低至 25 个高表达端粒酶的 HeLa 细胞。该策略为端粒酶活性的测定、癌细胞系与正常细胞系的区分以及端粒酶靶向抗癌药物的筛选提供了一个良好的平台。
