title: Dual immunosensor based on methylene blue-electroadsorbed graphene oxide for rapid detection of the influenza A virus antigen.
abstract: Rapid detection of influenza viral infections in poultry facilities is advantageous in several aspects such as environmental/personal safety, food-security, and socio-economy. Herein, we report the development of an electrochemical-based dual-sensor platform composed of methylene blue-electroadsorbed graphene oxide nanostructures modified with monoclonal antibodies against the HA proteins of H5N1 and H1N1. Bio-functional layers comprised of chitosan and protein-A molecules were implemented at the interface of the sensor element and antibodies, which synergistically enriched the bio-activity of immobilized antibodies for the immune complex formation. The differential pulse voltammetric signals resulted from the developed immunosensor platform exhibited a good correlation (R(2)=0.9978 for H1N1 and R(2)=0.9997 for H5N1) for the wide range of target concentrations 25-500pM). Chronoamperometric study also revealed the amplified current sensitivity of the immunoelectrodes even at the picomolar level. The proposed immunosensor design not only provides rapid analytical response time (<1min) but simplicity in fabrication and instrumentation, which paves an attractive platform for on-farm monitoring of viral infections.
authors.affiliation: BioNano Laboratory, School of Engineering, University of Guelph, Guelph, ON, Canada N1G 2W1.
authors.forename: Murugan
authors.initials: M
authors.lastname: Veerapandian
authors.affiliation: BioNano Laboratory, School of Engineering, University of Guelph, Guelph, ON, Canada N1G 2W1.
authors.forename: Robert
authors.initials: R
authors.lastname: Hunter
authors.affiliation: BioNano Laboratory, School of Engineering, University of Guelph, Guelph, ON, Canada N1G 2W1. Electronic address: sneethir@uoguelph.ca.
authors.forename: Suresh
authors.initials: S
authors.lastname: Neethirajan
mesh_terms: D000327:Adsorption; D000956:Antigens, Viral / Q000032:analysis* / Q000737:chemistry; D015374:Biosensing Techniques / Q000379:methods*; D004563:Electrochemistry; D006108:Graphite / Q000737:chemistry*; D007118:Immunoassay / Q000379:methods*; D053118:Influenza A Virus, H1N1 Subtype / Q000276:immunology; D053124:Influenza A Virus, H5N1 Subtype / Q000276:immunology; D008751:Methylene Blue / Q000737:chemistry*; D008958:Models, Molecular; D008968:Molecular Conformation; D010087:Oxides / Q000737:chemistry*; D013997:Time Factors
chemical_list: D000956:Antigens, Viral; D010087:Oxides; D006108:Graphite; D008751:Methylene Blue
keywords: Dual immunosensor; Electroadsorption; Graphene oxide; H1N1 flu detection; H5N1 flu detection; Methylene blue