Abstract
The COVID-19 pandemic is caused by SARS-CoV-2, a novel zoonotic coronavirus. Emerging evidence indicates that preexisting humoral immunity against other seasonal human coronaviruses (HCoVs) plays a critical role in the specific antibody response to SARS-CoV-2. However, current work to assess the effects of preexisting and cross-reactive anti-HCoVs antibodies has been limited. To address this issue, we have adapted our previously reported multiplex assay to simultaneously and quantitatively measure anti-HCoV antibodies. The full mPlex-CoV panel covers the spike (S) and nucleocapsid (N) proteins of three highly pathogenic HCoVs (SARS-CoV-1, SARS-CoV-2, MERS) and four human seasonal strains (OC43, HKU1, NL63, 229E). Combining this assay with volumetric absorptive microsampling (VAMS), we measured the anti-HCoV IgG, IgA, and IgM antibodies in fingerstick blood samples. The results demonstrate that the mPlex-CoV assay has high specificity and sensitivity. It can detect strain-specific anti-HCoV antibodies down to 0.1 ng/ml with 4 log assay range and with low intra- and inter-assay coefficients of variation (%CV). We also estimate multiple strain HCoVs IgG, IgA and IgM concentration in VAMS samples in three categories of subjects: pre-COVID-19 (n=21), post-COVID-19 convalescents (n=19), and COVID-19 vaccine recipients (n=14). Using metric multidimensional scaling (MDS) analysis, HCoVs IgG concentrations in fingerstick blood samples were well separated between the pre-COVID-19, post-COVID-19 convalescents, and COVID-19 vaccine recipients. In addition, we demonstrate how multi-dimensional scaling analysis can be used to visualize IgG mediated antibody immunity against multiple human coronaviruses. We conclude that the combination of VAMS and the mPlex-Cov assay is well suited to performing remote study sample collection under pandemic conditions to monitor HCoVs antibody responses in population studies.
Highlights
The coronavirus infectious disease 2019 (COVID-19) pandemic has taken over 2.5 million lives worldwide, and over 500,000 in the US, as of March 15, 2021 [1]
We describe the combination of volumetric absorptive microsampling (VAMS) and a 15antigen multiplex mPLEX-CoV assay for simultaneous measurement of IgG, IgA, and IgM against the S and N proteins of severe acute respiratory syndrome (SARS)-COV-1, SARS-CoV-2 and four seasonal human coronaviruses (HCoVs) (OC43, HKU1, NL63, 229E), as well as the S1, S2, and receptor-binding domain (RBD) protein domains of SARS-CoV-2
We adapted our multiplex antibody assay [18, 24] to detect antibodies that react to 12 HCoV S and N proteins, as well as antibodies that recognize the subunits of SARS-CoV-2, S1, S2 and RBD
Summary
The coronavirus infectious disease 2019 (COVID-19) pandemic has taken over 2.5 million lives worldwide, and over 500,000 in the US, as of March 15, 2021 [1]. We demonstrate a clear multidimensional separation between the pre-COVID, COVID, and COVID-vaccine groups in their IgG using metric multidimensional scaling (MDS) analysis of the measured IgG concentrations This proof-of-concept study demonstrates that fingerstick VAMS with the mPlex-CoV assay would significantly improve our ability to study the role of preexisting HCoV antibodies on SARS-CoV-2 infection and vaccination immune responses, and to conduct remote sample collection and multidimensional systems serology in COVID-19 population immunity studies and clinical trials. This will become increasingly important as newer SARS-CoV-2 variants emerge
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