Zinc Oxide Nanorod-Based Electrochemical Biosensors for Detecting SARS-CoV-2 Antibodies and Spike Protein
The COVID-19 pandemic exposed critical limitations in global diagnostic infrastructure and emphasized the need for rapid, decentralized, and accurate serological tools. In this study, we developed and evaluated a series of electrochemical biosensors based on zinc oxide nanorods (ZnONRs) for the detection of anti-SARS-CoV-2 antibodies and Spike protein antigens in human serum. Four biosensing platforms were constructed: (i) a device using the recombinant trimeric Spike protein from the SARS-CoV-2 wild-type (WT); (ii) an immunosensor functionalized with the Spike protein from the Gamma variant (P.1); (iii) a peptide-based biosensor employing the immunodominant B-cell epitope P44 (from the receptor-binding domain); and (iv) a biosensor using an ACE-2 peptide immobilized on Ni-doped ZnONRs for antigen detection.
All devices were fully characterized and validated with serum samples from infected, convalescent, vaccinated (CoronaVac, ChAdOx1-S, and BNT162b2), and pre-pandemic individuals. The WT Spike-based immunosensor demonstrated 88.67% sensitivity and 100% specificity, with a limit of detection (LOD) of 19.34 ng·mL⁻¹. The Gamma variant sensor showed an LOD of 52.55 ng·mL⁻¹ and revealed stronger antibody responses in BNT162b2-vaccinated individuals compared to ChAdOx1-S. The P44 peptide-based device effectively recognized WT antibodies targeting single-residue mutations (K417) in WT, Gamma, and Omicron variants, with sub-nanogram LODs. Finally, the ACE-2-based sensor enabled direct Spike antigen detection, with an LOD of 60.13 ng·mL⁻¹ and AUC of 0.917.
The results highlight the versatility, sensitivity, and diagnostic potential of the ZnONRs-based biosensors for point-of-care applications in immunological surveillance and COVID-19 variant monitoring.