A PCR-free colorimetric strategy for visualized assay of telomerase activity.
A PCR-free colorimetric strategy for visualized assay of telomerase activity.
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DOI：
10.1016/j.talanta.2017.09.070
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发表时间：
2018-02
期刊：
影响因子：
6.1
通讯作者：
T. Yu;Wei Zhao;Jingjuan Xu;Hongyuan Chen
中科院分区：
文献类型：
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作者：
T. Yu;Wei Zhao;Jingjuan Xu;Hongyuan Chen
这项工作报告了一种简单而强大的无聚合酶链反应（PCR）策略，用于可视化检测人类端粒酶活性。基于金纳米颗粒 (AuNP) 的比色策略与精心设计的酶辅助循环扩增相结合。简而言之，检测依赖于端粒酶诱导的端粒酶底物 (TS) 延长引物，该引物打开发夹 DNA 并与接头 DNA 杂交，从而触发 AuNP 聚集。传感系统中添加了切口核酸内切酶，杂交后切割接头DNA并释放互补链与接头DNA进行循环杂交，从而使端粒酶的检测具有高灵敏度。可以识别多达 25 个高表达端粒酶的 HeLa 细胞。该策略为端粒酶活性测定、癌细胞系与正常细胞系的分化以及端粒酶靶向抗癌药物的筛选提供了良好的平台。
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.