Multiparametric Integrated Molecular Detection of SARS-CoV-2 from Biofluids by Adapting Single Extracellular Vesicle Characterization Technologies

COVID-19, a global pandemic due to its rapid progression, is caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). There are different assays for COVID-19 testing, including nucleic acid, serological, antigen, and ancillary tests used in hospitals. In this study, a biochip assay that can simultaneously detect SARS-CoV-2 protein and RNA from biofluids by using surface markers, and simultaneously detect/quantify target proteins and RNA at single virus resolution was developed. Amplification of the signals were achieved by targeting different regions of the viral RNA (e.g., viral nucleocapsid N1, N2), and a tyramide signal amplification (TSA) assay for viral surface proteins. To demonstrate the clinical utility of the technology, it was deployed and tested using different biofluids, including NPS, and saliva from COVID-19 patients.