Analytical and clinical performances of five immunoassays for the detection of SARS-CoV-2 antibodies in comparison with neutralization activity

Abstract

Background: Reliable high-throughput serological assays for SARS-CoV-2 antibodies are urgently needed for the effective containment of the COVID-19 pandemic, as it is of crucial importance to understand the strength and duration of immunity after infection, and to make informed decisions concerning the activation or discontinuation of physical distancing restrictions.

Methods: In 184 serum samples from 130 COVID-19 patients and 54 SARS-CoV-2 negative subjects, the analytical and clinical performances of four commercially available chemiluminescent assays (Abbott SARS-Cov-2 IgG, Roche Elecsys anti-SARS-CoV-2, Ortho SARS-CoV-2 total and IgG) and one enzyme-linked immunosorbent assay (Diesse ENZY-WELL SARS-CoV-2 IgG) were evaluated and compared with the neutralization activity achieved using the plaque reduction neutralization test (PRNT).

Findings: Precision results ranged from 0.9% to 11.8% for all assays. Elecsys anti-SARS-CoV-2 demonstrated linearity of results at concentrations within the cut-off value. Overall, sensitivity ranged from 78.5 to 87.7%, and specificity, from 97.6 to 100%. On limiting the analysis to samples collected 12 days after onset of symptoms, the sensitivity of all assays increased, the highest value (95.2%) being obtained with VITRO Anti-SARS-CoV-2 Total and Architect SARS-CoV-2 IgG. The strongest PRNT₅₀ correlation with antibody levels was obtained with ENZY-Well SARS-CoV-2 IgG (R²adj = 0.569).

Interpretation: The results confirmed that all immunoassays had an excellent specificity, whereas sensitivity varied across immunoassays, depending strongly on the time interval between symptoms onset and sample collection. Further studies should be conducted to achieve a stronger correlation between antibody measurement and PRNT₅₀ in order to obtain useful information for providing a better management of COVID-19 patients, effective passive antibody therapy, and developing a vaccine against the SARS-CoV-2 virus.

Funding: None.

Keywords: Antibodies; Clinical performances; Immunoassays; SARS-CoV-2; Serology.

Fig. 1

Linearity assessment of the studies immunoassays.

Fig. 2

Comparison of plaque reduction neutralization test (PRNT) and immunoassay results. A) Roche Elecsys Anti-SARS-CoV-2 Total against N antigen, B) Abbott Architect Anti-SARS-CoV-2 IgG against N antigen, C) Liaison Diasorin Anti-SARS-CoV-2 IgG Against S1/S2 protein, D) Ortho Clinical Diagnostics VITROS Anti-SARS-CoV-2 IgG against S protein, E) Ortho Clinical Diagnostics VITROS Anti-SARS-CoV-2 Total against S1 protein, F) Enzy-Well, Anti-SARS-CoV-2 IgG against native antigen; G) ENZY-WELL Anti-SARS-CoV-2 IgA against native antigen, H) ENZY-WELL Anti-SARS-CoV-2 IgM against native antigen. For all the comparisons n = 68 samples were evaluated (except for Liaison Diasorin Anti-SARS-CoV-2 IgG Against S1/S2 protein where n = 52). For Liaison Diasorin, reagents were available for 52 out of the 68 tested neutralization samples.

Fig. 3

Plaque reduction neutralization test (PRNT) results, disease severity and time from symptoms onset (days). Asympt/Pauci = asymptomatic/paucisymptomatic SARS-CoV-2 patients; Moderate and Severe = SARS-CoV-2 positive patients without and with air ventilation support, respectively; M (within bars) = males; F (within bars) = females.