Ag(I)-coordinated hairpin DNA for homogenous electronic monitoring of hepatitis C virus accompanying isothermal cycling signal amplification strategy
Ag(I)-coordinated hairpin DNA for homogenous electronic monitoring of hepatitis C virus accompanying isothermal cycling signal amplification strategy
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Ag(I) 协调的发夹 DNA 伴随等温循环信号放大策略用于丙型肝炎病毒同质电子监测
DOI：
10.1016/j.bios.2015.05.058
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发表时间：
2015
影响因子：
12.6
通讯作者：
Wang Youmei
中科院分区：
文献类型：
--
作者：
Lu Minghua;Xu Linfang;Zhang Xiaona;Xiao Rui;Wang Youmei
本研究通过胞嘧啶- Ag+ -胞嘧啶配位化学，设计了一种新的均质电子监测平台，用于在无固定化Ag(I)辅助发夹DNA上灵敏检测丙型肝炎病毒（HCV）。该实验由靶诱导的Ag+从发夹DNA解离和等温环状链位移聚合（ICSDP）反应组成。引入目标分析物后，HCV DNA首先与发夹DNA杂交，破坏Ag(I)配位发夹探针并释放配位Ag+离子，然后新形成的DNA双链在引物和聚合酶的作用下诱导ICSDP反应，然后位移的靶DNA重新触发Ag(I)配位发夹DNA与靶再循环，从而在检测细胞内形成大量的游离Ag+离子。释放的Ag+离子可以很容易地被带负电的丝网印刷碳电极捕获，随后对捕获的Ag+离子进行阳极剥离伏安检测，形成阳极电流，用于在所施加电位内产生电化学信号。在最佳条件下，基于icsdp的均相传感系统可用于低至2.3 pM浓度的HCV DNA检测。同一批次的内、间变异系数分别小于9.5%和10.5%。对5份临床血清标本的分析表明，所开发的方法与商用Cobas®Amplicor HCV测试分析仪的结果吻合良好
This work designs a new homogenous electronic monitoring platform for sensitive detection of hepatitis C virus (HCV) on an immobilization-free Ag(I)-assisted hairpin DNA through the cytosine–Ag+–cytosine coordination chemistry. The assay consists of target-induced Ag+dissociation from hairpin DNA and an isothermal circular strand-displacement polymerization (ICSDP) reaction. Upon target analyte introduction, HCV DNA initially hybridizes with hairpin DNA to disrupt the Ag(I)-coordinated hairpin probe and releases the coordinated Ag+ion, then the newly formed DNA duplex induces the ICSDP reaction with the aid of primer and polymerase, and then the displaced target DNA retriggers Ag(I)-coordinated hairpin DNA with target recycling, thereby resulting in formation of numerous free Ag+ions in the detection cell. The released Ag+ions can be readily captured by the negatively charged screen-printed carbon electrode, and subsequent anodic-stripping voltammetric detection of the captured Ag+ions are conducted to form the anodic current for the production of the electrochemical signal within the applied potential. Under optimal conditions, the ICSDP-based homogenous sensing system can be utilized for the detection of HCV DNA at a concentration as low as 2.3 pM. Intra- and inter-assay coefficients of variation with identical batches are below 9.5% and 10.5%, respectively. The analysis in 5 clinical serum specimens shows good accordance between results obtained by the developed method and commercial Cobas® Amplicor HCV Test Analyzer