MICA: Community Based Point-of-Need Integrated Diagnostics for SARS-CoV-2

Currently, validated diagnostic tests for SARS-CoV-2 infections include laboratory-based tests involving Reverse Transcriptase (RT) Polymerase Chain Reaction (PCR) or RT-isothermal Loop-Mediated Amplification (RT-LAMP), using swabs from the upper respiratory tract, as well as blood-based IgM/IgG/IgA enzyme-linked immunosorbent assays (ELISAs). Rapid diagnostic tests (RDTs) lateral flow immunodiagnostic devices (LFD) are also available for community testing. Most recently DNA testing ex-laboratories has been achieved, albeit with complex and expensive units (e.g. DNAnudge). In general, although these commercial products provide solutions for testing in many scenarios, there remains a need for low-cost, point-of-need nucleic acid sensors, able to detect SARS-CoV-2 infections in a timely manner, providing multiplexed tests (e.g. with RSV, influenza), without the need for expensive equipment. We have developed a novel SARS-CoV-2 LAMP assay, targeting sequences in the ORF1 ab region and N gene simultaneously (as recommended by China CDC and USA FDA). The assay has been optimised, now giving a limit of detection <64 copies/ml and validated clinically against 7 other respiratory viruses, showing no cross reactivity. We propose to incorporate this into a multiplexed community-use, hand-held nucleic acid-based sensor as a self-contained single-use disposable device. A nasopharyngeal swab or saliva sample will be placed into an injection-molded cartridge, containing all reagents to perform sample enrichment and RT-LAMP. The multiplexed result is read visually on an integrated paper-LFD. The device enables (i) sample enrichment, reducing the elution volume from a swab and (ii) simple methods for thermal management, and will be validated in a statistically-powered study, to ensure the test's rapid translation to our clinical and industrial partners.