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Microscopy-based assay for semi-quantitative detection of SARS-CoV-2 specific antibodies in human sera

Constantin Pape, Roman Remme, Adrian Wolny, Sylvia Olberg, Steffen Wolf, Lorenzo Cerrone, Mirko Cortese, Severina Klaus, Bojana Lucic, Stephanie Ullrich, Maria Anders-Össwein, Stefanie Wolf, Cerikan Berati, Christopher Neufeld, Markus Ganter, Paul Schnitzler, Uta Merle, Marina Lusic, Steeve Boulant, Megan Stanifer, Ralf Bartenschlager, Fred A. Hamprecht, Anna Kreshuk, Christian Tischer, Hans-Georg Kräusslich, Barbara Müller, Vibor Laketa
doi: https://doi.org/10.1101/2020.06.15.152587
Constantin Pape
1HCI/IWR, Heidelberg University, Heidelberg, Germany
2European Molecular Biology Laboratory, Heidelberg, Germany
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Roman Remme
1HCI/IWR, Heidelberg University, Heidelberg, Germany
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Adrian Wolny
1HCI/IWR, Heidelberg University, Heidelberg, Germany
2European Molecular Biology Laboratory, Heidelberg, Germany
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Sylvia Olberg
3Department of Infectious Diseases, Virology, University Hospital Heidelberg, Heidelberg, Germany
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Steffen Wolf
1HCI/IWR, Heidelberg University, Heidelberg, Germany
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Lorenzo Cerrone
1HCI/IWR, Heidelberg University, Heidelberg, Germany
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Mirko Cortese
4Department of Infectious Diseases, Molecular Virology, University Hospital Heidelberg, Heidelberg, Germany
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Severina Klaus
5Department of Infectious Diseases, Parasitology, University Hospital Heidelberg, Heidelberg, Germany
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Bojana Lucic
6Department of Infectious Diseases, Integrative Virology, University Hospital Heidelberg, Heidelberg, Germany
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Stephanie Ullrich
3Department of Infectious Diseases, Virology, University Hospital Heidelberg, Heidelberg, Germany
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Maria Anders-Össwein
3Department of Infectious Diseases, Virology, University Hospital Heidelberg, Heidelberg, Germany
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Stefanie Wolf
3Department of Infectious Diseases, Virology, University Hospital Heidelberg, Heidelberg, Germany
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Cerikan Berati
4Department of Infectious Diseases, Molecular Virology, University Hospital Heidelberg, Heidelberg, Germany
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Christopher Neufeld
4Department of Infectious Diseases, Molecular Virology, University Hospital Heidelberg, Heidelberg, Germany
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Markus Ganter
5Department of Infectious Diseases, Parasitology, University Hospital Heidelberg, Heidelberg, Germany
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Paul Schnitzler
3Department of Infectious Diseases, Virology, University Hospital Heidelberg, Heidelberg, Germany
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Uta Merle
7Department of Gastroenterology and Hepatology, University Hospital of Heidelberg, Heidelberg, Germany
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Marina Lusic
6Department of Infectious Diseases, Integrative Virology, University Hospital Heidelberg, Heidelberg, Germany
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Steeve Boulant
3Department of Infectious Diseases, Virology, University Hospital Heidelberg, Heidelberg, Germany
8Research Group “Cellular polarity and viral infection”, German Cancer Research Center (DKFZ), Heidelberg, Germany
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Megan Stanifer
4Department of Infectious Diseases, Molecular Virology, University Hospital Heidelberg, Heidelberg, Germany
8Research Group “Cellular polarity and viral infection”, German Cancer Research Center (DKFZ), Heidelberg, Germany
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Ralf Bartenschlager
4Department of Infectious Diseases, Molecular Virology, University Hospital Heidelberg, Heidelberg, Germany
9German Center for Infection Research, Heidelberg, Germany
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Fred A. Hamprecht
1HCI/IWR, Heidelberg University, Heidelberg, Germany
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Anna Kreshuk
2European Molecular Biology Laboratory, Heidelberg, Germany
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Christian Tischer
2European Molecular Biology Laboratory, Heidelberg, Germany
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Hans-Georg Kräusslich
3Department of Infectious Diseases, Virology, University Hospital Heidelberg, Heidelberg, Germany
9German Center for Infection Research, Heidelberg, Germany
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Barbara Müller
3Department of Infectious Diseases, Virology, University Hospital Heidelberg, Heidelberg, Germany
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  • For correspondence: vibor.laketa@med.uni-heidelberg.de barbara.mueller@med.uni-heidelberg.de
Vibor Laketa
3Department of Infectious Diseases, Virology, University Hospital Heidelberg, Heidelberg, Germany
9German Center for Infection Research, Heidelberg, Germany
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  • For correspondence: vibor.laketa@med.uni-heidelberg.de barbara.mueller@med.uni-heidelberg.de
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Abstract

Emergence of the novel pathogenic coronavirus Sars-CoV-2 and its rapid pandemic spread presents numerous questions and challenges that demand immediate attention. Among these is the urgent need for a better understanding of humoral immune response against the virus and assessment of seroprevalence levels in the population, both of which form the basis for developing public health strategies to control viral spread. For this sensitive, specific and quantitative serological assays are required. Here, we describe the development of a semi-quantitative high-content microscopy-based assay for detection of three major classes (IgG, IgA and IgM) of Sars-CoV-2 specific antibodies in human samples. The possibility to detect antibodies against the entire viral proteome together with a robust semi-automated image analysis workflow resulted in improvement of sensitivity and specificity compared to an approved ELISA-based diagnostic test. Combining both resulted in maximum specificity in a negative control cohort, while maintaining high sensitivity. The procedure described here is compatible with high-throughput microscopy approaches and may be applied for serological analysis of other virus infections.

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Microscopy-based assay for semi-quantitative detection of SARS-CoV-2 specific antibodies in human sera
Constantin Pape, Roman Remme, Adrian Wolny, Sylvia Olberg, Steffen Wolf, Lorenzo Cerrone, Mirko Cortese, Severina Klaus, Bojana Lucic, Stephanie Ullrich, Maria Anders-Össwein, Stefanie Wolf, Cerikan Berati, Christopher Neufeld, Markus Ganter, Paul Schnitzler, Uta Merle, Marina Lusic, Steeve Boulant, Megan Stanifer, Ralf Bartenschlager, Fred A. Hamprecht, Anna Kreshuk, Christian Tischer, Hans-Georg Kräusslich, Barbara Müller, Vibor Laketa
bioRxiv 2020.06.15.152587; doi: https://doi.org/10.1101/2020.06.15.152587
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Microscopy-based assay for semi-quantitative detection of SARS-CoV-2 specific antibodies in human sera
Constantin Pape, Roman Remme, Adrian Wolny, Sylvia Olberg, Steffen Wolf, Lorenzo Cerrone, Mirko Cortese, Severina Klaus, Bojana Lucic, Stephanie Ullrich, Maria Anders-Össwein, Stefanie Wolf, Cerikan Berati, Christopher Neufeld, Markus Ganter, Paul Schnitzler, Uta Merle, Marina Lusic, Steeve Boulant, Megan Stanifer, Ralf Bartenschlager, Fred A. Hamprecht, Anna Kreshuk, Christian Tischer, Hans-Georg Kräusslich, Barbara Müller, Vibor Laketa
bioRxiv 2020.06.15.152587; doi: https://doi.org/10.1101/2020.06.15.152587

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