Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Suzhou, 215163, People's Republic of China.
University of Science and Technology of China, Hefei, 230026, People's Republic of China.
Mikrochim Acta. 2018 Aug 1;185(8):398. doi: 10.1007/s00604-018-2936-x.
The paper describes a voltammetric method for the quantitation of the activity of telomerase extracted from cancer cells. A thiolated single-stranded telomerase substrate primer was firstly immobilized on a gold electrode. In the presence of a mixture of telomerase and deoxynucleotide triphosphates, the primer becomes elongated and contains repetitive nucleotide sequences (TTAGGG). After hybridization with blocker DNA, gold nanoparticles are added and captured by the elongated single-stranded DNA. This reduces the charge transfer resistance of the gold electrode. The telomerase activity is then quantified via differential pulse voltammetry, typically at 0.12 V (vs. SCE). The method is PCR-free, rapid, and convenient. It was applied to the detection of HeLa cells via the telomerase activity of lysed cells. The detection range was from 500 to 50,000 cells/mL and the detection limit was as low as 500 cells/mL. Graphical abstract A telomerase substrate (TS) primer is immobilized on a gold electrode as the sensing interface to detect the activity of telomerase extracted from cancer cells. Unmodified gold nanoparticles (AuNPs) are utilized which change the electrochemical responses.
本文描述了一种用于定量测定从癌细胞中提取的端粒酶活性的伏安法。首先将巯基化的单链端粒酶底物引物固定在金电极上。在端粒酶和脱氧核苷酸三磷酸混合物存在的情况下，引物延伸并包含重复的核苷酸序列（TTAGGG）。与封闭 DNA 杂交后，加入金纳米粒子并被延伸的单链 DNA 捕获。这降低了金电极的电荷转移电阻。然后通过差分脉冲伏安法（通常在 0.12 V（相对于 SCE））定量端粒酶活性。该方法无 PCR、快速且方便。它通过裂解细胞中端粒酶的活性被应用于 HeLa 细胞的检测。检测范围为 500 至 50,000 个细胞/mL，检测限低至 500 个细胞/mL。 图表摘要 将端粒酶底物（TS）引物固定在金电极上作为传感界面，以检测从癌细胞中提取的端粒酶的活性。未修饰的金纳米粒子（AuNPs）被利用，这改变了电化学响应。
