A label-free and signal-on electrochemical aptasensor for ultrasensitive kanamycin detection based on exonuclease recycling cleavage
A label-free and signal-on electrochemical aptasensor for ultrasensitive kanamycin detection based on exonuclease recycling cleavage
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一种基于核酸外切酶回收切割的无标记、信号电化学适体传感器，用于超灵敏卡那霉素检测
DOI：
10.1039/c5ay02754c
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发表时间：
2016-01
影响因子：
3.1
通讯作者：
Chenhe Lu
中科院分区：
文献类型：
--
作者：
Yuanyuan Xu;Linghao Sun;Xiaocui Huang;Yangyang Sun;Chenhe Lu
利用核酸外切酶 III (Exo III) 的优先外脱氧核糖核酸酶活性，结合适体的高度特异性识别，基于适体触发的 Exo III 回收裂解，提出了一种用于超灵敏卡那霉素 (Kan) 检测的无标记和信号电化学适体传感器。带有 5' 硫醇基团的适体互补 DNA (cDNA) 自组装到金电极表面，用于随后与卡那霉素特异性适体 (K-适体) 杂交。形成的双链体 DNA 具有 3' 羟基平端作为识别元件，并且 cDNA 可以被 Exo III 逐步去除，最终导致 K-适配体的释放，用于连续新的杂交和切割循环。检测到分析物后，Kan 和 K-适体的结合阻碍了 Exo III 催化的切割过程，并且保留的 cDNA 与 Kan 的浓度成正比，通过 RuHex 的吸附对 Kan 的浓度进行电化学定量。由于电化学方法的高灵敏度和 Exo III 的循环信号放大，所提出的适体传感器可以检测低至 1 pM 水平的 Kan。更重要的是，它是无标记、信号开启和表面束缚的，这使得该生物传感器更适合和稳定地应用于实际样品中的Kan检测。
Taking advantage of the preferential exodeoxyribonuclease activity of exonuclease III (Exo III) in combination with the highly specific recognition of aptamers, a label-free and signal-on electrochemical aptasensor for ultrasensitive kanamycin (Kan) detection has been proposed based on aptamer triggered Exo III recycling cleavage. Aptamer complementary DNA (cDNA) with a 5′ thiol group is self-assembled onto a gold electrode surface for the following hybridization with a kanamycin specific aptamer (K-aptamer). The formed duplex DNA has 3′ hydroxyl blunt termini as a recognition element, and cDNA could be stepwise removed by Exo III ultimately resulting in the release of the K-aptamer for successive new cycles of hybridization and cleavage. Upon sensing the analyte, the combination of Kan and K-aptamer impedes the cleavage process catalyzed by Exo III, and the retained cDNA is proportional to the concentration of Kan, which is electrochemically quantified via the adsorption of RuHex. Due to the high sensitivity of the electrochemical method and the recycling signal amplification of Exo III, the proposed aptasensor can detect Kan down to a 1 pM level. More importantly, it is label-free, signal-on and surface-tethered, which make this biosensor more suitable and stable for the application of Kan detection in real samples.
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