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Comparative Study
. 2021 May;40(5):1063-1071.
doi: 10.1007/s10096-021-04169-7. Epub 2021 Feb 3.

Sensitivity of anti-SARS-CoV-2 serological assays in a high-prevalence setting

Lisa Müller #  1 , Philipp Niklas Ostermann #  1 , Andreas Walker  1 , Tobias Wienemann  2 , Alexander Mertens  3 , Ortwin Adams  1 , Marcel Andree  1 , Sandra Hauka  1 , Nadine Lübke  1 , Verena Keitel  3 , Ingo Drexler  1 , Veronica Di Cristanziano  4 , Derik Franz Hermsen  5 , Rolf Kaiser  4 , Friedrich Boege  5 , Florian Klein  4 , Heiner Schaal  1 , Jörg Timm  6 , Tina Senff  7
Affiliations
Comparative Study

Sensitivity of anti-SARS-CoV-2 serological assays in a high-prevalence setting

Lisa Müller et al. Eur J Clin Microbiol Infect Dis. 2021 May.

Abstract

Evaluation and power of seroprevalence studies depend on the performed serological assays. The aim of this study was to assess four commercial serological tests from EUROIMMUN, DiaSorin, Abbott, and Roche as well as an in-house immunofluorescence and neutralization test for their capability to identify SARS-CoV-2 seropositive individuals in a high-prevalence setting. Therefore, 42 social and working contacts of a German super-spreader were tested. Consistent with a high-prevalence setting, 26 of 42 were SARS-CoV-2 seropositive by neutralization test (NT), and immunofluorescence test (IFT) confirmed 23 of these 26 positive test results (NT 61.9% and IFT 54.8% seroprevalence). Four commercial assays detected anti-SARS-CoV-2 antibodies in 33.3-40.5% individuals. Besides an overall discrepancy between the NT and the commercial assays regarding their sensitivity, this study revealed that commercial SARS-CoV-2 spike-based assays are better to predict the neutralization titer than nucleoprotein-based assays are.

Keywords: COVID-19; Immunofluorescence test; Neutralizing antibodies; SARS-CoV-2; Serology; Seroprevalence.

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Conflict of interest statement

The authors declare no competing interests.

Figures

Fig. 1
Fig. 1
SARS-CoV-2-neutralizing antibodies stratified according to status of study participants and exemplary immunofluorescence test results, Heinsberg District, Germany, April 2020 (n = 42). FITC: fluorescein isothiocyanate; NT: neutralization test. a Neutralization test results of 42 individuals grouped by their status in PCR confirmed (red), symptomatic (blue), and asymptomatic (black) and 11 control sera from healthy individuals sampled before December 2019. The reciprocal of the NT titer is depicted, and bars represent the respective median. The cut-off was defined as ≥20. One-way ANOVA was used to compare groups (**p ≤ 0.01 and ***p ≤ 0.001). b Exemplary anti-SARS-CoV-2 IgG immunofluorescence test results of 3 out of 42 tested individuals. Phase contrast (a−e) and FITC fluorescence detected at 488 nm (f−j). Serum of a severe hospitalized COVID-19 case served as a positive control (a+f) and (b+g) depict the result of a negative control serum. IFT results from a patient with a high NT titer (c+h; NT titer 10,240), a low NT titer (d+i NT titer 40), and no neutralization potential (e+j). Scale bar is 100 μm
Fig. 2
Fig. 2
Correlation between commercial SARS-CoV-2 antibody tests and the neutralization titer, Heinsberg District, Germany, April 2020 (n = 42). AU: arbitrary units; COI: cut-off index; EI: EUROIMMUN; N: nucleocapsid; NT: neutralization test; OD: optical density; r: correlation coefficient; S1: spike domain 1; S2: spike domain 2; S/C: sample/control; SARS-CoV-2: severe acute respiratory syndrome coronavirus 2. The reciprocal of the NT titer is depicted. RT-PCR-confirmed SARS-CoV-2 infections are depicted in red and symptomatic individuals in blue. All asymptomatic individuals are displayed in black. a and b EUROIMMUN-anti-SARS-CoV-2 IgA and IgG ELISA (Euroimmun). c LIAISON® SARS-CoV-2 S1/S2 IgG (DiaSorin). d SARS-CoV-2 IgG CMIA (Abbott). e Elecsys® anti-SARS-CoV-2 ECLIA test (Roche). The dotted lines indicate the cut-off values recommended by the respective manufacturer to determine positive and negative test results. The borderline area if applicable is indicated in yellow and the vertical line represents the positive cut-off of an NT titer ≥20
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