Journal
VIRUSES-BASEL
Volume 13, Issue 2, Pages -Publisher
MDPI
DOI: 10.3390/v13020143
Keywords
SARS-CoV-2; serology; serodiagnosis; TR-FRET; immunoassay
Categories
Funding
- Sigrid Juselius Foundation
- Magnus Ehrnrooth Foundation
- Finnish Society of Sciences and Letters
- Finska Lakaresallskapet
- Finnish Medical Foundation
- Academy of Finland [1308613, 1336490, 336439, 335527]
- Juho Vainio Foundation
- Jane and Aatos Erkko Foundation
- European Union Horizon 2020 programme VEO (Versatile emerging infectious disease observatory) [874735]
- University of Helsinki
- Research Funds of University of Helsinki [TYH 2018322]
- Helsinki University Hospital [TYH 2018322]
- Academy of Finland (AKA) [336439, 335527] Funding Source: Academy of Finland (AKA)
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This study demonstrates the applicability of LFRET in detecting SARS-CoV-2 antibodies, showing high sensitivity and specificity for antibodies against SP and NP.
Accurate and rapid diagnostic tools are needed for management of the ongoing coronavirus disease 2019 (COVID-19) pandemic. Antibody tests enable detection of individuals past the initial phase of infection and help examine vaccine responses. The major targets of human antibody response in severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are the spike glycoprotein (SP) and nucleocapsid protein (NP). We have developed a rapid homogenous approach for antibody detection termed LFRET (protein L-based time-resolved Forster resonance energy transfer immunoassay). In LFRET, fluorophore-labeled protein L and antigen are brought to close proximity by antigen-specific patient immunoglobulins of any isotype, resulting in TR-FRET signal. We set up LFRET assays for antibodies against SP and NP and evaluated their diagnostic performance using a panel of 77 serum/plasma samples from 44 individuals with COVID-19 and 52 negative controls. Moreover, using a previously described SP and a novel NP construct, we set up enzyme linked immunosorbent assays (ELISAs) for antibodies against SARS-CoV-2 SP and NP. We then compared the LFRET assays with these ELISAs and with a SARS-CoV-2 microneutralization test (MNT). We found the LFRET assays to parallel ELISAs in sensitivity (90-95% vs. 90-100%) and specificity (100% vs. 94-100%). In identifying individuals with or without a detectable neutralizing antibody response, LFRET outperformed ELISA in specificity (91-96% vs. 82-87%), while demonstrating an equal sensitivity (98%). In conclusion, this study demonstrates the applicability of LFRET, a 10-min mix and read assay, to detection of SARS-CoV-2 antibodies.
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