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Dual detection of COVID-19 antigens and antibodies using nanoscale fluorescent plasmonic substrates.

There is a continuing need for biosensors that can be used in the diagnosis of COVID-19 infection and to measure a subject's immune response to the virus itself (SARS-CoV-2). In this study, grating-coupled fluorescent plasmonic (GC-FP)-based detection of SARS-CoV-2 antigens was coupled with antibody detection to yield a dual-mode detection assay. Pairs of capture and detection antibodies were screened for direct detection of the SARS-CoV-2 nucleocapsid (Nuc) antigen, which were then combined with an existing GC-FP antibody detection assay. Nuc could be detected as low as 1 µg/mL concentrations, while antibodies were detectable to 50 ng/mL. The dual detection assay was tested by adding purified Nuc antigen to serum from a polymerase chain reaction (PCR)-positive COVID-19-infected individual. Using this sample, co-detection of Nuc antigen and anti-spike protein antibodies was successfully performed on a single GC-FP chip. Total assay time was 1 h, making this the first known example of rapid dual antibody and antigen detection on the same biosensor chip.