The analytical specificity and microbial interference of a SARS-CoV-2 biosensor detection platform are elucidated. A cost-effective and highly sensitive detection system for the virus has been developed with the capability of producing quantitative results comparable with polymerase chain reaction (PCR) within 30 seconds. This could meet the demand for a fast diagnosis solution needed for the ongoing global pandemic. Disposable strips were biofunctionalized and immobilized with monoclonal SARS-CoV-2 antibodies. A printed circuit board embedded with a metal–oxide–semiconductor field-effect transistor (MOSFET) was also designed. The strips were connected to the gate electrode of the MOSFET, which received a synchronous pulse along with the drain electrode. The resulting waveform from the drain was then converted to digital readouts corresponding to virus or spike protein concentrations. We investigated 26 common organisms which are likely presented in the respiratory system along with 5 pathogens from the same genetic family of the SARS-CoV-2 virus for having cross-reaction or microbial interference, either of which would hinder the efficacy of the system. None of these organisms decreased the virus detection effectiveness of the sensor system.