Characterization of a Pan-Immunoglobulin Assay Quantifying Antibodies Directed against the Receptor Binding Domain of the SARS-CoV-2 S1-Subunit of the Spike Protein: A Population-Based Study

Abstract

Pan-immunoglobulin assays can simultaneously detect IgG, IgM and IgA directed against the receptor binding domain (RBD) of the S1 subunit of the spike protein (S) of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2 S1-RBD Ig). In this work, we aim to evaluate a quantitative SARS-CoV-2 S1-RBD Ig electrochemiluminescence immunoassay (ECLIA) regarding analytical, diagnostic, operational and clinical characteristics. Our work takes the form of a population-based study in the principality of Liechtenstein, including 125 cases with clinically well-described and laboratory confirmed SARS-CoV-2 infection and 1159 individuals without evidence of coronavirus disease 2019 (COVID-19). SARS-CoV-2 cases were tested for antibodies in sera taken with a median of 48 days (interquartile range, IQR, 43–52) and 139 days (IQR, 129–144) after symptom onset. Sera were also tested with other assays targeting antibodies against non-RBD-S1 and -S1/S2 epitopes. Sensitivity was 97.6% (95% confidence interval, CI, 93.2–99.1), whereas specificity was 99.8% (95% CI, 99.4–99.9). Antibody levels linearly decreased from hospitalized patients to symptomatic outpatients and SARS-CoV-2 infection without symptoms (p < 0.001). Among cases with SARS-CoV-2 infection, smokers had lower antibody levels than non-smokers (p = 0.04), and patients with fever had higher antibody levels than patients without fever (p = 0.001). Pan-SARS-CoV-2 S1-RBD Ig in SARS-CoV-2 infection cases significantly increased from first to second follow-up (p < 0.001). A substantial proportion of individuals without evidence of past SARS-CoV-2 infection displayed non-S1-RBD antibody reactivities (248/1159, i.e., 21.4%, 95% CI, 19.1–23.4). In conclusion, a quantitative SARS-CoV-2 S1-RBD Ig assay offers favorable and sustained assay characteristics allowing the determination of quantitative associations between clinical characteristics (e.g., disease severity, smoking or fever) and antibody levels. The assay could also help to identify individuals with antibodies of non-S1-RBD specificity with potential clinical cross-reactivity to SARS-CoV-2.

Persistence and baseline determinants of seropositivity in health care workers up to nine months after COVID-19

“…Lower antibody levels in nurses and physicians than other HCW could indicate a lower exposure due to PPE use and higher awareness of risks 14 . Smoking had previously been associated with lower antibody responses 15 and we show that this effect persists after several months primarily affecting IgA, the main mucosal antibody.…”

Persistence and baseline determinants of seropositivity and reinfection rates in health care workers up to 12.5 months after COVID-19

“…Lower antibody levels in nurses and physicians than other HCW could indicate a lower exposure due to PPE use and higher awareness of risks [19]. Smoking had previously been associated with lower antibody responses [22,23] and we show that this effect persists after several months primarily affecting IgA, the main mucosal antibody.…”

Evaluation of a Broad Panel of SARS-CoV-2 Serological Tests for Diagnostic Use

“…Recently, a number of studies have compared and validated smaller panels ( n = 1–7) of SARS-CoV-2 serological tests for diagnostic use [ 9 , 11 , 12 , 13 , 14 , 15 , 16 , 17 , 18 , 19 , 20 , 21 , 22 , 23 , 24 , 25 ]. In the present study, we intended to systematically cross-compare a large number of SARS-CoV-2 serological tests, and thus analyzed a broad panel of SARS-CoV-2 ECLIA-, ELISA-, and LFA-based tests which were available in May 2020, using a relatively large number of sera from patients with COVID-19, and from controls.…”