SARS-CoV-2-Specific Antibody Detection for Seroepidemiology: A Multiplex Analysis Approach Accounting for Accurate Seroprevalence

• Published in: The Journal of Infectious Diseases Vol. 222; no. 9; pp. 1452 - 1461
• Main Authors: den Hartog, Gerco, Schepp, Rutger M, Kuijer, Marjan, GeurtsvanKessel, Corine, van Beek, Josine, Rots, Nynke, Koopmans, Marion P G, van der Klis, Fiona R M, van Binnendijk, Robert S
• Format: Journal Article Web Resource
• Language: English
• Published: United States Oxford University Press 01.10.2020
• Subjects: Adaptive Immunity; Adult; Aged; Aged, 80 and over; Antibodies, Viral - blood; Area Under Curve; Betacoronavirus - immunology; Case-Control Studies; Coronavirus Infections - blood; Coronavirus Infections - epidemiology; COVID-19; Female; Humans; Immunoassay; Immunoglobulin G - blood; Major and Brief Reports; Male; Middle Aged; Netherlands - epidemiology; Nuclear Proteins - immunology; Pandemics; Patient Acuity; Pneumonia, Viral - blood; Pneumonia, Viral - epidemiology; ROC Curve; SARS-CoV-2; Seroconversion; Seroepidemiologic Studies; Spike Glycoprotein, Coronavirus - immunology; Netherlands; COVID-19; endemic coronavirus; influenza-like Illness (ILI); RBD; IgG; multiplex bead-based immune assay; nucleoprotein; specificity; spike S1; sensitivity
• ISSN: 0022-1899; 1537-6613
• DOI: 10.1093/infdis/jiaa479

The COVID-19 pandemic necessitates better understanding of the kinetics of antibody production induced by infection with SARS-CoV-2. We aimed to develop a high-throughput multiplex assay to detect antibodies to SARS-CoV-2 to assess immunity to the virus in the general population. Spike protein subunits S1 and receptor binding domain, and nucleoprotein were coupled to microspheres. Sera collected before emergence of SARS-CoV-2 (n = 224) and of non-SARS-CoV-2 influenza-like illness (n = 184), and laboratory-confirmed cases of SARS-CoV-2 infection (n = 115) with various severities of COVID-19 were tested for SARS-CoV-2-specific IgG concentrations. Our assay discriminated SARS-CoV-2-induced antibodies and those induced by other viruses. The assay specificity was 95.1%-99.0% with sensitivity 83.6%-95.7%. By merging the test results for all 3 antigens a specificity of 100% was achieved with a sensitivity of at least 90%. Hospitalized COVID-19 patients developed higher IgG concentrations and the rate of IgG production increased faster compared to nonhospitalized cases. The bead-based serological assay for quantitation of SARS-CoV-2-specific antibodies proved to be robust and can be conducted in many laboratories. We demonstrated that testing of antibodies against multiple antigens increases sensitivity and specificity compared to single-antigen-specific IgG determination.