There is tremendous interest in development of systems for fast and high throughput testing technology for transmissive diseases and emergency medicine, especially after the outbreak of SARS-CoV-2 virus (COVID-19). To date (Apr 23rd, 2021), the number of infected populations worldwide has reached above 145 millions with more than 3 millions fatalities from the pandemic. Electrochemical sensing could potentially meet the demand of rapid yet low-cost detection and diagnosis for decentralized and widespread use. Recent advances in electrochemical sensing for COVID-19 include analog signal detection using graphene based field effect transistors (FET) [1], electrochemical impedance-based detection [2], as well as sensing of the N-gene of SARS-CoV-2 using antisense oligonucleotides [3]. In this work, we present a modular and low-cost platform Si-MOSFET based biosensor system utilizing disposable cartilages for sensing of SARS-CoV-2 spiked peptide and inactivated virus.Figure 1 shows the overall schematics of the sensing system, which consists of a Si enhancement mode MOSFET (Texas Instruments SN74S124N) based circuit board and biofunctionalized test strips. The sensing test strips, Figure 2, are connected between the VGG and the gate electrode. A voltage-controlled oscillator (VCO), shown in Fig 3, converted the output drain waveform into a digital reading. The starting materials for the sensing cartridges are multi-electrode graphite-based sensor strips with a single electrode plated with Au for sensing. Figure 4 shows the surface morphology of the electroplated surface. The Au-plated surface was then functionalized, subsequently, with thioglycolic acid (TGA), N, N’-dicyclohexylcarbodi-imide and N-hydroxysuccinimide, to form chemical bonding between Au surface and the desired antibody. Diluted antibody was then incubated on the Au electrode surface to finalize the biofunctionalization.Figure 5 shows a schematic of the synchronous drain (VDD) and gate (VGG) waveform, as well as the resultant dynamic drain current (ID/Isen) and voltage (VD/Vsen) caused by the electric field induced deformation of the antigen/antibody complex [4]. Figure 6 shows the dynamic drain waveform of the NMOS transistor at different tested spike peptide concentrations. Figure 7 shows the voltage extracted at 750μs of the drain waveform, which is designated as the analog signal reported in this work. Two types of antibodies, SARS-CoV-Spike ECD monoclonal antibody and polyclonal spike antibody, were tested in this study. The built-in digital readout allows direct readout of the relative virus concentration. Both antibodies show sensitivity in range of 10-15 to 10-5 g/mL with digital signal sensitivity of 53/dec for monoclonal antibody and 56/dec for polyclonal antibody. Figure 8 shows the digital outputs for human inactivated virus (ATCC VR-1986HK) under test. The sensor strips with electrodes presented in Figure 2 can reach detection limits down to 100 PFU/mL with digital sensitivity of 150/dec for monoclonal antibody and 454/dec for polyclonal antibody. By slightly modifying the sensor tip configuration, the sensitivity for inactivated virus detection can be lowered to 5 PFU/mL. For the same range of peptide/virus concentration tested, a larger change in drain voltage/digital reading represents a high sensitivity to doped solution due to the different epitopes these antibodies interact with. The fast response time and low detection of limit exemplified by this technique provides an opportunity for inexpensive semiconductor-based sensor systems in such applications. This approach is versatile and can be functionalized with different antibodies for different viruses and the cartridges printed with multiple functional areas to test for multiple variants.[1] G. Seo et al., ACS Nano, vol. 14, p. 5135 (2020)[2] M. Z. Rashed et al., Biosens. Bioelectron., vol. 171, p. 112709 (2021)[3] M. Alafeef, et al, ACS Nano, vol. 14, pp. 17028–17045 (2020)[4] S. Shan et al., IEEE Trans. Biomed. Circuits Syst. vol 14, 1362 (2020) Figure 1
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