Screen-Printed Graphene/Carbon Electrodes on Paper Substrates as Impedance Sensors for Detection of Coronavirus in Nasopharyngeal Fluid Samples

Severe acute respiratory syndrome (SARS-CoV-2), the causative agent of the global pandemic, which has resulted in more than one million deaths with tens of millions reported cases, requires a fast, accurate, and portable testing mechanism operable in the field environment. Electrochemical sensors, based on paper substrates with portable electrochemical devices, can prove an excellent alternative in mitigating the economic and public health effects of the disease. Herein, we present an impedance biosensor for the detection of the SARS-CoV-2 spike protein utilizing the IgG anti-SARS-CoV-2 spike antibody. This label-free platform utilizing screen-printed electrodes works on the principle of redox reaction impedance of a probe and can detect antigen spikes directly in nasopharyngeal fluid as well as virus samples collected in the universal transport medium (UTM). High conductivity graphene/carbon ink is used for this purpose so as to have a small background impedance that leads to a wider dynamic range of detection. Antibody immobilization onto the electrode surface was conducted through a chemical entity or a biological entity to see their effect; where a biological immobilization can enhance the antibody loading and thereby the sensitivity. In both cases, we were able to have a very low limit of quantification (i.e., 0.25 fg/mL), however, the linear range was 3 orders of magnitude wider for the biological entity-based immobilization. The specificity of the sensor was also tested against high concentrations of H1N1 flu antigens with no appreciable response. The most optimized sensors are used to identify negative and positive COVID-19 samples with great accuracy and precision.