Direct bioelectronic detection of SARS-CoV-2 from saliva using single-molecule field-effect transistor array

Direct bioelectronic detection of SARS-CoV-2 from saliva using single-molecule field-effect transistor array Nucleic acid tests have become the gold-standard for diagnostic testing for COVID-19, usually performedin specialized laboratories. Most are based on reverse-transcription quantitative polymerase chain reaction(qRT-PCR). The time required for specimen transport and processing results in a turnaround time that is typi-cally several days. The few rapid (<1 hour) point-of-care (POC) tests are more expensive, still require samplepreparation and specialized reagents, and do not have the throughput needed for population surveillance. Di-rect testing for the virus, which also reduces requirements for multiple reagents, is a necessary step to improv-ing diagnostic testing. While four such antigen tests have been approved for detection of SARS-CoV-2 basedon immunoassays to the N protein, sensitivity is limited and no quantitation of viral load is possible. We will address this gap by using DiagnostikosTM, an in-development rapid POC platform for direct, real-time, multiplexed, quantitative bioelectronic detection of biomolecules that employs an all-electronic detectiondevice that functions at the single-molecule level. These single-molecule field-effect transistors (smFETs) arearrayed on a complementary metal-oxide-semiconductor (CMOS) integrated circuit chip. Chips will interfacewith an envisioned USB-stick-form-factor reader device. Robust single-domain antibodies, known as nanobod-ies and immobilized on these devices, are used for sensitive detection of viral particles and viral debris. Theuse of multiple nanobodies for a single protein and nanobodies for different proteins in a single assay allowsfor significant improvements in specificity. Nanobodies will be specific for one or more of the four major struc-tural proteins in SARS-CoV-2; the nucleocapsid (N) protein engulfing the viral RNA, the spike (S) protein, themembrane (M) protein and the envelope (E) protein. No sample preparation or specialized reagents are re-quired for detection, and the device will be designed to operate with saliva, which has very recently beenshown to be a reliable medium for detecting SARS-CoV-2. Individual sensor chips can be manufactured at acost of $35. With the addition of other nanobodies, these large dense arrays can also allow detection of manypathogens in a single test. In this Direct-To-Phase-2 SBIR program we will pursue several key innovations that are required to makesuch a platform possible, including isolation of nanobodies for key structure proteins of SARS-CoV-2 (SpecificAim 1), development of the smFET platform for antigen detection (Specific Aim 2), development of largeCMOS arrays of these smFET devices (Specific Aim 3), and verification of detection in increasingly complexsamples up to and including clinical samples (Specific Aim 4). This project is a partnership between universityresearchers who developed the smFET technology and a venture-based start-up venture, Quicksilver Biosci-ences, spun out to commercialize smFET technology and develop smFET/CMOS arrays for molecular diag-nostic applications.