A highly specific and sensitive serological assay detects SARS-CoV-2 antibody levels in COVID-19 patients that correlate
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ORIGINAL PAPER
A highly specific and sensitive serological assay detects SARS‑CoV‑2 antibody levels in COVID‑19 patients that correlate with neutralization David Peterhoff1 · Vivian Glück2 · Matthias Vogel2 · Philipp Schuster1 · Anja Schütz1 · Philip Neubert2 · Veruschka Albert2 · Stefanie Frisch2 · Mara Kiessling2 · Philip Pervan2 · Maren Werner1 · Nicole Ritter2 · Leon Babl2 · Maria Deichner2 · Frank Hanses3,4 · Matthias Lubnow5 · Thomas Müller5 · Dirk Lunz6 · Florian Hitzenbichler3 · Franz Audebert7 · Viola Hähnel8 · Robert Offner8 · Martina Müller9 · Stephan Schmid9 · Ralph Burkhardt10 · Thomas Glück11 · Michael Koller12 · Hans Helmut Niller1 · Bernhard Graf6 · Bernd Salzberger3 · Jürgen J. Wenzel2 · Jonathan Jantsch1,2 · André Gessner1,2 · Barbara Schmidt1,2 · Ralf Wagner1,2 Received: 1 July 2020 / Accepted: 7 August 2020 © The Author(s) 2020
Abstract Objective The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic challenges national health systems and the global economy. Monitoring of infection rates and seroprevalence can guide public health measures to combat the pandemic. This depends on reliable tests on active and former infections. Here, we set out to develop and validate a specific and sensitive enzyme linked immunosorbent assay (ELISA) for detection of anti-SARS-CoV-2 antibody levels. Methods In our ELISA, we used SARS-CoV-2 receptor-binding domain (RBD) and a stabilized version of the spike (S) ectodomain as antigens. We assessed sera from patients infected with seasonal coronaviruses, SARS-CoV-2 and controls. We determined and monitored IgM-, IgA- and IgG-antibody responses towards these antigens. In addition, for a panel of 22 sera, virus neutralization and ELISA parameters were measured and correlated. Results The RBD-based ELISA detected SARS-CoV-2-directed antibodies, did not cross-react with seasonal coronavirus antibodies and correlated with virus neutralization (R2 = 0.89). Seroconversion started at 5 days after symptom onset and led to robust antibody levels at 10 days after symptom onset. We demonstrate high specificity (99.3%; N = 1000) and sensitivity (92% for IgA, 96% for IgG and 98% for IgM; > 10 days after PCR-proven infection; N = 53) in serum. Conclusions With the described RBD-based ELISA protocol, we provide a reliable test for seroepidemiological surveys. Due to high specificity and strong correlation with virus neutralization, the RBD ELISA holds great potential to become a preferred tool to assess thresholds of protective immunity after infection and vaccination. Keywords SARS-CoV-2 · COVID-19 · Antibody test · ELISA · Serology · Virus neutralization · Assay validation · Spike protein · S protein · Receptor binding domain
Introduction Barbara Schmidt and Ralf Wagner contributed equally. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s15010-020-01503-7) contains supplementary material, which is available to authorized users. * David Peterhoff [email protected] * Barbara Schmidt barbara.schmidt@uk
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