title: Ag(I)-coordinated hairpin DNA for homogenous electronic monitoring of hepatitis C virus accompanying isothermal cycling signal amplification strategy.
abstract: 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.
authors.affiliation: Institute of Environmental and Analytical Science, School of Chemistry and Chemical Engineering, Henan University, Kaifeng 475004, Henan, PR China. Electronic address: minghualu88@yeah.net.
authors.forename: Minghua
authors.initials: M
authors.lastname: Lu
authors.affiliation: Institute of Environmental and Analytical Science, School of Chemistry and Chemical Engineering, Henan University, Kaifeng 475004, Henan, PR China.
authors.forename: Linfang
authors.initials: L
authors.lastname: Xu
authors.affiliation: Institute of Environmental and Analytical Science, School of Chemistry and Chemical Engineering, Henan University, Kaifeng 475004, Henan, PR China.
authors.forename: Xiaona
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Institute of Environmental and Analytical Science, School of Chemistry and Chemical Engineering, Henan University, Kaifeng 475004, Henan, PR China.
authors.forename: Rui
authors.initials: R
authors.lastname: Xiao
authors.affiliation: Institute of Environmental and Analytical Science, School of Chemistry and Chemical Engineering, Henan University, Kaifeng 475004, Henan, PR China.
authors.forename: Youmei
authors.initials: Y
authors.lastname: Wang
mesh_terms: D015374:Biosensing Techniques / Q000379:methods*; D015342:DNA Probes / Q000737:chemistry / Q000235:genetics; D055664:Electrochemical Techniques / Q000379:methods; D016174:Hepacivirus / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D055029:Inverted Repeat Sequences; D007454:Iodides; D012367:RNA, Viral / Q000097:blood / Q000235:genetics / Q000302:isolation & purification; D015203:Reproducibility of Results; D018030:Silver Compounds
chemical_list: D015342:DNA Probes; D007454:Iodides; D012367:RNA, Viral; D018030:Silver Compounds; C030584:silver iodide
keywords: Ag(I)-coordinated hairpin DNA; Electrochemical DNA sensor; Hepatitis C virus; Homogeneous assay; Isothermal circular strand-displacement polymerization