Sensitive electrochemiluminescent detection of telomerase activity based on nicking enzyme assisted signal amplification

检出限 电化学发光 端粒酶 生物传感器 环介导等温扩增 滚动圆复制 寡核苷酸 化学 适体 互补DNA 分子生物学 DNA 色谱法 生物化学 生物 DNA聚合酶 基因
作者
Shu-Ying Ye,Chang-Gang Pan,Yu-Hua Dai,Guoxi Liang
出处
期刊:Microchemical Journal [Elsevier]
卷期号:165: 106123-106123 被引量:8
标识
DOI:10.1016/j.microc.2021.106123
摘要

Herein, an ultrasensitive biosensor for the detection of telomerase activity was constructed by utilizing [email protected] dots nanohybrids ([email protected]) as a high efficiency electrochemiluminescence (ECL) emitter and nicking enzyme assisted signal amplification (NESA) technology. Firstly, the self-enhanced [email protected] was synthesized from the reduction of HAuCl4 with carbon dots (CDs), which served as reductant and stabilizer. The prepared hybrids exhibited significant enhancement in the ECL emission compared to the single CDs. Enhanced sensitivity was thus achieved by using these [email protected] as ECL labels. Next, an isothermal amplification with nicking enzyme (Nb.BbvcI)-assisted recycle was employed to convert a small amount of telomerase to numerous output signal DNA (sDNA). The obtained sDNA was then linked with [email protected] and immobilized on a capture DNA (cDNA), and connected to the working electrode through oligonucleotide hybridization, resulting in a quantitative ECL reading. As a consequence, by employment of the dual signal amplification strategy, the proposed biosensor exhibited excellent sensitivity for telomerase activity detection with a detection limit down to 1 HeLa cell, which is comparable or even better than the most reported methods. The method also displayed a wide linear response ranges and good reproducibility. This work not only expanded the application of high-efficiency ECL indicators [email protected], but also offered a novel PCR-free ECL sensing platform for detecting telomerase activity that is helpful for real-time monitoring, point-of-care testing and on-site analysis.
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