电化学发光法通过杂交链式反应和 G-四链体/血红素 DNA zyme 生物传感策略灵敏检测不同癌细胞中的甲基转移酶活性。
An electrochemiluminescence assay for sensitive detection of methyltransferase activity in different cancer cells by hybridization chain reaction coupled with a G-quadruplex/hemin DNAzyme biosensing strategy.
机构信息
Jiangsu Key Laboratory of New Power Batteries, Jiangsu Collaborative Innovation Center of Biomedical Functional Materials, Jiangsu Key Laboratory of Biomedical Materials, National and Local Joint Engineering Research Center of Biomedical Functional Materials, College of Chemistry and Materials Science, Nanjing Normal University, Nanjing 210097, P. R. China.
出版信息
Analyst. 2017 May 30;142(11):2013-2019. doi: 10.1039/c7an00486a.
In this work, a highly sensitive electrochemiluminescence (ECL) assay was fabricated for the detection of human DNA (cytosine-5)-methyltransferase1 (DNMT1) activity in cancer cells. The ECL assay coupled hybridization chain reaction with a G-quadruplex/hemin DNAzyme biosensing strategy. The ECL intensity changes (ΔI) allowed detection of DNMT1 activity down to 0.09 U mL, and ΔI was proportional to the logarithm of the activity of DNMT1 within the range of 1.0 to 30.0 U mL in buffer solution. It also showed high sensitivity to DNMT1 activity in A549 cells, with a detection limit of about 2 cells. This ECL assay provides a promising platform for profiling of the mutational cells of tumors and shows a great potential for application to DNA methylation-related clinical diagnostics.
在这项工作中，我们构建了一种高灵敏度的电化学发光（ECL）分析方法，用于检测癌细胞中人类 DNA（胞嘧啶-5）-甲基转移酶 1（DNMT1）的活性。该 ECL 分析方法将杂交链式反应与 G-四链体/血红素 DNA 酶生物传感策略相结合。ECL 强度变化（ΔI）可检测到低至 0.09 U mL 的 DNMT1 活性，并且 ΔI 与 DNMT1 活性的对数成正比，在缓冲溶液中 DNMT1 活性的范围为 1.0 至 30.0 U mL。它还显示出对 A549 细胞中 DNMT1 活性的高灵敏度，检测限约为 2 个细胞。该 ECL 分析方法为肿瘤突变细胞的分析提供了一个有前途的平台，并显示出在与 DNA 甲基化相关的临床诊断中应用的巨大潜力。
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