SPCE-Based Electrochemical Immunosensor for Influenza A (H1) Detection in Serum and Nasopharyngeal Samples

Acute respiratory diseases caused by viral pathogens such as Influenza A continue to represent a major global health challenge, emphasizing the need for rapid, sensitive, and accessible diagnostic tools. In this work, a carbon screen-printed electrode (SPCE)-based electrochemical immunosensor for the detection of an Influenza A (H1) antigen is reported, incorporating a comparative electrochemical evaluation of four electrode materials. Fe₃O₄ nanoparticles, Fe₃O₄@C nanoparticles, graphene quantum dots (GQDs), and gold nanoparticles (AuNPs) were systematically assessed by cyclic voltammetry to evaluate their electrocatalytic performance. The highest electrochemical response was selected for biosensor construction. The immunosensor was fabricated by immobilizing antibodies on a modified SPCE and characterized using differential pulse voltammetry (DPV). A concentration-dependent response was observed for H1 antigen concentrations ranging from 0 to 300 ng/mL, with a minimum detectable concentration (MDC) of 1 ng/mL and limit of detection (LOD) of 176 ng/mL and 45 ng/mL for serum and nasopharyngeal swabs, respectively. The biosensor performance was specifically evaluated in complex biological fluids, demonstrating reproducible performance and moderate selectivity against non-target influenza subtypes. Overall, this study highlights the critical role of electrode material selection in determining electrochemical immunosensor performance and supports the potential of SPCE-based platforms for the screening of an Influenza A (H1) antigen in point-of-care-oriented applications.