title: Graphene quantums dots combined with endonuclease cleavage and bidentate chelation for highly sensitive electrochemiluminescent DNA biosensing.
abstract: A novel strategy for highly sensitive electrochemiluminescence (ECL) detection of DNA was proposed based on site-specific cleavage of BamHI endonuclease combined with the excellent ECL activity of graphene quantum dots (GQDs) and bidentate chelation of the dithiocarbamate DNA (DTC-DNA) probe assembly. The difference between photoluminescence and ECL spectral peaks suggested that a negligible defect existed on the GQDs surface for generation of an ECL signal. The formed DTC-DNA was directly attached to the gold surface by bidentate anchoring (S-Au-S bonds), which conferred a strong affinity between the ligands and the gold surface, increasing the robustness of DNA immobilization on the gold surface. BamHI endonuclease site-specifically recognized and cleaved the duplex symmetrical sequence, which made the double-stranded DNA fragments and GQDs break off from the electrode surface, inducing a decrease of the ECL signal. Using hepatitis C virus-1b genotype complementary DNA (HCV-1b cDNA) as a model, a novel signal-off ECL DNA biosensor was developed based on variation of the ECL intensity before and after digestion of the DNA hybrid. Electrochemical impedance spectroscopy confirmed the successful fabrication of the ECL DNA biosensor. This ECL biosensor for HCV-1b cDNA determination exhibited a linear range from 5 fM to 100 pM with a detection limit of 0.45 fM at a signal-to-noise ratio of 3 and showed satisfactory selectivity and good stability, which validated the feasibility of the designed strategy. The proposed strategy may be conveniently combined with other specific biological recognition events for expansion of the biosensing application, especially in clinical diagnoses.
authors.affiliation: Jiangsu Collaborative Innovation Center of Biomedical Functional Materials and Jiangsu Key Laboratory of Biofunctional Materials, College of Chemistry and Materials Science, Nanjing Normal University , Nanjing, 210023, P. R. China.
authors.forename: Jing
authors.initials: J
authors.lastname: Lou
authors.forename: Shanshan
authors.initials: S
authors.lastname: Liu
authors.forename: Wenwen
authors.initials: W
authors.lastname: Tu
authors.forename: Zhihui
authors.initials: Z
authors.lastname: Dai
mesh_terms: D015374:Biosensing Techniques / Q000379:methods*; D002614:Chelating Agents / Q000378:metabolism; D018076:DNA, Complementary / Q000235:genetics; D004279:DNA, Viral / Q000032:analysis* / Q000235:genetics; D015245:Deoxyribonuclease BamHI / Q000378:metabolism*; D055664:Electrochemical Techniques / Q000379:methods*; D006108:Graphite / Q000737:chemistry*; D016174:Hepacivirus / Q000235:genetics*; D006526:Hepatitis C / Q000175:diagnosis / Q000235:genetics / Q000821:virology; D006801:Humans; D057230:Limit of Detection; D008163:Luminescent Measurements / Q000379:methods*; D045663:Quantum Dots; D059629:Signal-To-Noise Ratio; D013859:Thiocarbamates / Q000737:chemistry
chemical_list: D002614:Chelating Agents; D018076:DNA, Complementary; D004279:DNA, Viral; D013859:Thiocarbamates; D006108:Graphite; D015245:Deoxyribonuclease BamHI