• Infection · Feb 2021

    A highly specific and sensitive serological assay detects SARS-CoV-2 antibody levels in COVID-19 patients that correlate with neutralization.

    • David Peterhoff, Vivian Glück, Matthias Vogel, Philipp Schuster, Anja Schütz, Philip Neubert, Veruschka Albert, Stefanie Frisch, Mara Kiessling, Philip Pervan, Maren Werner, Nicole Ritter, Leon Babl, Maria Deichner, Frank Hanses, Matthias Lubnow, Thomas Müller, Dirk Lunz, Florian Hitzenbichler, Franz Audebert, Viola Hähnel, Robert Offner, Martina Müller, Stephan Schmid, Ralph Burkhardt, Thomas Glück, Michael Koller, Hans Helmut Niller, Bernhard Graf, Bernd Salzberger, Jürgen J Wenzel, Jonathan Jantsch, André Gessner, Barbara Schmidt, and Ralf Wagner.
    • Institute for Medical Microbiology and Hygiene, University of Regensburg, Regensburg, Germany. david.peterhoff@ur.de.
    • Infection. 2021 Feb 1; 49 (1): 75-82.

    ObjectiveThe 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.MethodsIn 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.ResultsThe 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.ConclusionsWith 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.

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