Rapid response flow cytometric assay for the detection of antibody responses to SARS-CoV-2
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ORIGINAL ARTICLE
Rapid response flow cytometric assay for the detection of antibody responses to SARS-CoV-2 Dennis Lapuente 1 & Clara Maier 1 & Pascal Irrgang 1 & Julian Hübner 1 & Antonia Sophia Peter 1 & Markus Hoffmann 2 & Armin Ensser 1 & Katharina Ziegler 3 & Thomas H. Winkler 4 & Torsten Birkholz 5 & Andreas E. Kremer 6 & Philipp Steininger 1 & Klaus Korn 1 & Frank Neipel 1 & Klaus Überla 1 & Matthias Tenbusch 1 Received: 28 May 2020 / Accepted: 9 October 2020 # The Author(s) 2020
Abstract SARS-CoV-2 has emerged as a previously unknown zoonotic coronavirus that spread worldwide causing a serious pandemic. While reliable nucleic acid–based diagnostic assays were rapidly available, only a limited number of validated serological assays were available in the early phase of the pandemic. Here, we evaluated a novel flow cytometric approach to assess spike-specific antibody responses.HEK 293T cells expressing SARS-CoV-2 spike protein in its natural confirmation on the surface were used to detect specific IgG and IgM antibody responses in patient sera by flow cytometry. A soluble angiotensin-converting-enzyme 2 (ACE-2) variant was developed as external standard to quantify spike-specific antibody responses on different assay platforms. Analyses of 201 pre-COVID-19 sera proved a high assay specificity in comparison to commercially available CLIA and ELISA systems, while also revealing the highest sensitivity in specimens from PCR-confirmed SARS-CoV-2-infected patients. The external standard allowed robust quantification of antibody responses among different assay platforms. In conclusion, our newly established flow cytometric assay allows sensitive and quantitative detection of SARS-CoV-2-specific antibodies, which can be easily adopted in different laboratories and does not rely on external supply of assay kits. The flow cytometric assay also provides a blueprint for rapid development of serological tests to other emerging viral infections Keywords Coronavirus . SARS-CoV-2 . Serology . Antibodies . Flow cytometry
Introduction * Matthias Tenbusch [email protected] 1
Institute of Clinical and Molecular Virology, University Hospital Erlangen, Friedrich-Alexander University Erlangen-Nürnberg, Erlangen, Germany
2
Infection Biology Unit, German Primate Center-Leibniz Institute for Primate Research, Göttingen, Germany
3
Institute of Clinical Hygiene, Medical Microbiology and Infectiology, Paracelsus Medical University, Nürnberg, Germany
4
Department of Biology, Division of Genetics, Nikolaus-Fiebiger-Center for Molecular Medicine, Friedrich-Alexander University Erlangen-Nürnberg, Erlangen, Germany
5
Department of Anaesthesiology, University Hospital Erlangen, Erlangen, Germany
6
Department of Medicine 1, Gastroenterology, Pneumology and Endocrinology, University Hospital Erlangen, Friedrich-Alexander University Erlangen-Nürnberg, Erlangen, Germany
In early December 2019, a novel zoonotic coronavirus (CoV) caused a cluster of pneumonia cases in Wuhan, China [1]. Since then, the virus has spread globally
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