Rapid testing for CoV-2 RNA in swabs

clinical trial - Rapid and extensive testing of both personnel and patients is crucial for proper management SARS-CoV-2 infections and decision-making in times of the current pandemic outbreak. However, point-of-care (POC) testing in emergency units or outpatient clinics is not possible yet at a larger scale using established PCR-based methods. Therefore, we tested two PCR-independent methods to detect viral RNA from primary material (swabs) circumventing the need of RNA preparation. Firstly, SHERLOCK, developed by Feng Zhang at MIT(1), uses Recombinase Polymerase Amplification (RPA) combined with Cas13a-dependent detection of RNA as a second step followed by a lateral-flow assay on a test strip. As a simple alternative, loop-mediated amplification (LAMP) only requires one incubation step with detection based either on a colorimetric assay or lateral flow strips. SHERLOCK and LAMP can be performed in 60 or 30 minutes, respectively. Based on our preliminary data, we can show that both methods work on isolated viral RNA as well as on direct material from pharyngeal swabs - a crucial pre-requisite for point-of-care testing. However, both approaches still lack sensitivity as compared to standard qPCR testing. In this proposal we aim to (1) improve and combine aspects of both SHERLOCK and LAMP to develop an easy and fast diagnostic method allowing for viral RNA detection without RNA isolation at a sensitivity comparable to qPCR and (2) to test these approaches in the setting of an emergency room.