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Rapid response flow cytometric assay for the detection of antibody responses to SARS-CoV-2.

Authors
  • Lapuente, Dennis1
  • Maier, Clara1
  • Irrgang, Pascal1
  • Hübner, Julian1
  • Peter, Antonia Sophia1
  • Hoffmann, Markus2
  • Ensser, Armin1
  • Ziegler, Katharina3
  • Winkler, Thomas H4
  • Birkholz, Torsten5
  • Kremer, Andreas E6
  • Steininger, Philipp1
  • Korn, Klaus1
  • Neipel, Frank1
  • Überla, Klaus1
  • Tenbusch, Matthias7
  • 1 Institute of Clinical and Molecular Virology, University Hospital Erlangen, Friedrich-Alexander University Erlangen-Nürnberg, Erlangen, Germany. , (Germany)
  • 2 Infection Biology Unit, German Primate Center-Leibniz Institute for Primate Research, Göttingen, Germany. , (Germany)
  • 3 Institute of Clinical Hygiene, Medical Microbiology and Infectiology, Paracelsus Medical University, Nürnberg, Germany. , (Germany)
  • 4 Department of Biology, Division of Genetics, Nikolaus-Fiebiger-Center for Molecular Medicine, Friedrich-Alexander University Erlangen-Nürnberg, Erlangen, Germany. , (Germany)
  • 5 Department of Anaesthesiology, University Hospital Erlangen, Erlangen, Germany. , (Germany)
  • 6 Department of Medicine 1, Gastroenterology, Pneumology and Endocrinology, University Hospital Erlangen, Friedrich-Alexander University Erlangen-Nürnberg, Erlangen, Germany. , (Germany)
  • 7 Institute of Clinical and Molecular Virology, University Hospital Erlangen, Friedrich-Alexander University Erlangen-Nürnberg, Erlangen, Germany. [email protected] , (Germany)
Type
Published Article
Journal
European Journal of Clinical Microbiology & Infectious Diseases
Publisher
Springer-Verlag
Publication Date
Oct 20, 2020
Identifiers
DOI: 10.1007/s10096-020-04072-7
PMID: 33078221
Source
Medline
Keywords
Language
English
License
Unknown

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.

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