Journal
BIOSENSORS & BIOELECTRONICS
Volume 73, Issue -, Pages 195-201Publisher
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2015.05.058
Keywords
Hepatitis C virus; Homogeneous assay; Electrochemical DNA sensor; Ag(l)-coordinated hairpin DNA; Isothermal circular strand-displacement polymerization
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Funding
- National Natural Science Foundation of China [21305029, 21477033]
- High-Level Talent from Henan University [B2012156]
- State Key Laboratory of the Cotton Biology Open Fund [CB2013A22]
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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 polyrnerase, 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 (R) Amplicor HCV Test Analyzer (C) 2015 Elsevier B.V. All rights reserved.
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