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We present a highly sensitive and selective nano-biosensor for rapid, stable and highly reproducible detection of ascorbic acid (AA) in the presence of dopamine, uric acid and other interferences by a three-layer sandwich arrangement of nitrogen-doped functionalized graphene (NFG), silver nanoparticles (AgNPs) and nanostructured polyaniline (PANI) nanocomposite. The enhanced AA electrochemical properties of the NFG/AgNPs/PANI electrode is attributed to the superior conductivity of the NFG-PANI and the excellent catalytic activity of AgNPs. The critical modification of the AgNPs-grafted NFG-PANI coated on very low-cost fluorine doped tin oxide electrode (FTOE) increased the charge transfer conductivity of the electrode (the resistance drops down from 11,000 Ω to 6 Ω). The nano-biosensor was used to accurately detect AA in vitamin C tablets with the recovery of 98%. The sensor demonstrated a low detection limit of 8 µM (S/N = 3) with a very wide linear detection range of 10–11,460 µM, good reproducibility and excellent selectivity performance for AA detection. The results demonstrate that this nanocomposite is a promising candidate for rapid and selective detection of AA in practical clinical samples.

The widespread and long-lasting effect of the COVID-19 pandemic has called attention to the significance of technological advances in the rapid diagnosis of SARS-CoV-2 virus. This study reports the use of a highly stable buffer-based zinc oxide/reduced graphene oxide (bbZnO/rGO) nanocomposite coated on carbon screen-printed electrodes for electrochemical immuno-biosensing of SARS-CoV-2 nuelocapsid (N-) protein antigens in spiked and clinical samples. The incorporation of a salt-based (ionic) matrix for uniform dispersion of the nanomixture eliminates multistep nanomaterial synthesis on the surface of the electrode and enables a stable single-step sensor nanocoating. The immuno-biosensor provides a limit of detection of 21 fg/mL over a linear range of 1–10 000 pg/mL and exhibits a sensitivity of 32.07 ohms·mL/pg·mm2 for detection of N-protein in spiked samples. The N-protein biosensor is successful in discriminating positive and negative clinical samples within 15 min, demonstrating its proof of concept used as a COVID-19 rapid antigen test.

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Multiplex electrochemical biosensors have been used for eliminating the matrix effect in complex bodily fluids or enabling the detection of two or more bioanalytes, overall resulting in more sensitive assays and accurate diagnostics. Many electrochemical biosensors lack reliable and low-cost multiplexing to meet the requirements of point-of-care detection due to either limited functional biosensors for multi-electrode detection or incompatible readout systems. We developed a new dual electrochemical biosensing unit accompanied by a customized potentiostat to address the unmet need for point-of-care multi-electrode electrochemical biosensing. The two-working electrode system was developed using screen-printing of a carboxyl-rich nanomaterial containing ink, with both working electrodes offering active sites for recognition of bioanalytes. The low-cost bi-potentiostat system (∼$80) was developed and customized specifically to the bi-electrode design and used for rapid, repeatable, and accurate measurement of electrochemical impedance spectroscopy signals from the dual biosensor. This binary electrochemical data acquisition (Bi-ECDAQ) system accurately and selectively detected SARS-CoV-2 Nucleocapsid protein (N-protein) in both spiked samples and clinical nasopharyngeal swab samples of COVID-19 patients within 30 min. The two working electrodes offered the limit of detection of 116 fg/mL and 150 fg/mL, respectively, with the dynamic detection range of 1–10,000 pg/mL and the sensitivity range of 2744–2936 Ω mL/pg.mm 2 for the detection of N-protein. The potentiostat performed comparable or better than commercial Autolab potentiostats while it is significantly lower cost. The open-source Bi-ECDAQ presents a customizable and flexible approach towards addressing the need for rapid and accurate point-of-care electrochemical biosensors for the rapid detection of various diseases.

Optimized self-contained microfluidic platform allows for single-step detection of proteins, through passive delivery of the dry-stored reagents required for immunosensing. Novel autonomous performance of the platform advances the point-of-care utilization of electrochemical protein sensing.