Label-Free Detection of Human Coronaviruses in Infected Cells Using Enhanced Darkfield Hyperspectral Microscopy (EDHM)

Human coronaviruses (HCoV) are causative agents of mild to severe intestinal and respiratory infections in humans. In the last 15 years, we have witnessed the emergence of three zoonotic, highly pathogenic HCoVs. Thus, early, and accurate detection of these viral pathogens is not only essential for preventing transmission but also for the timely treatment and monitoring of drug resistance. Herein, we applied enhanced darkfield hyperspectral microscopy (EDHM), a novel non-invasive, label-free diagnostic tool for rapid and accurate identification of two strains of HCoVs, i.e., OC43 and 229E. The EDHM technology allows collecting the optical image with both spectral and spatial details in a single measurement without direct contact between the specimen and the sensor. Thus, it can provide the direct mapping of spectral signatures specific for a given viral strain in a complex biological milieu. Our study demonstrated distinct spectral patterns for HCoV-OC43 and HCoV-229E virions in the solution, which can serve as distinguishable parameters for their differentiation. Furthermore, spectral signatures for both HCoV strains in the infected cells displayed a considerable peak wavelength shift compared to the uninfected cell samples indicating that the EDHM is applicable to detect and differentiate between HCoV infected and uninfected cells.