Verena Krähling1, Sandro Halwe2, Cornelius Rohde2, Dirk Becker2, Susanne Berghöfer3, Christine Dahlke4, Markus Eickmann3, Meryem S Ercanoglu5, Lutz Gieselmann5, Astrid Herwig3, Alexandra Kupke2, Helena Müller3, Petra Neubauer-Rädel3, Florian Klein6, Christian Keller7, Stephan Becker2. 1. Institute of Virology, Philipps University Marburg, Marburg, Germany; German Center for Infection Research (DZIF), Partner Site Gießen-Marburg-Langen, Marburg, Germany. Electronic address: kraehliv@staff.uni-marburg.de. 2. Institute of Virology, Philipps University Marburg, Marburg, Germany; German Center for Infection Research (DZIF), Partner Site Gießen-Marburg-Langen, Marburg, Germany. 3. Institute of Virology, Philipps University Marburg, Marburg, Germany. 4. First Department of Medicine, Division of Infectious Diseases, University Medical Center Hamburg-Eppendorf, Hamburg, Germany; Department for Clinical Immunology of Infectious Diseases, Bernhard Nocht Institute for Tropical Medicine, Hamburg, Germany; German Center for Infection Research, Hamburg-Lübeck-Borstel-Riems, Germany. 5. Laboratory of Experimental Immunology, Institute of Virology, Faculty of Medicine and University Hospital of Cologne, University of Cologne, Cologne, Germany. 6. Laboratory of Experimental Immunology, Institute of Virology, Faculty of Medicine and University Hospital of Cologne, University of Cologne, Cologne, Germany; German Center for Infection Research (DZIF), Partner Site Bonn-Cologne, Cologne, Germany. 7. Institute of Virology, Philipps University Marburg, Marburg, Germany; Institute of Virology, University Hospital Giessen and Marburg, Marburg, Germany.
Abstract
The current Severe acute respiratory syndrome-related coronavirus 2 (SARS-CoV-2) pandemic is a public health emergency of international concern. Sensitive and precise diagnostic tools are urgently needed. In this study, we developed a SARS-CoV-2 spike (S1) protein enzyme-linked immunosorbent assay (ELISA) to detect SARS-CoV-2-specific antibodies. The SARS-CoV-2 S1 ELISA was found to be specific [97.8% (95% CI, 96.7% - 98.5%)], reproducible and precise (intra-assay coefficient of variability (CV) 5.3%, inter-assay CV 7.9%). A standard curve and the interpolation of arbitrary ELISA units per milliliter served to reduce the variability between different tests and operators. Cross-reactivity to other human coronaviruses was addressed by using sera positive for MERS-CoV- and hCoV HKU1-specific antibodies. Monitoring antibody development in various samples of twenty-three and single samples of twenty-nine coronavirus disease 2019 (COVID-19) patients revealed seroconversion and neutralizing antibodies against authentic SARS-CoV-2 in all cases. The comparison of the SARS-CoV-2 (S1) ELISA with a commercially available assay showed a better sensitivity for the in-house ELISA. The results demonstrate a high reproducibility, specificity and sensitivity of the newly developed ELISA, which is suitable for the detection of SARS-CoV-2 S1 protein-specific antibody responses.
The current Severe acute respiratory syndrome-related coronavirus 2 (n>an class="Species">SARS-CoV-2) pandemic is a public health emergency of international concern. Sensitive and precise diagnostic tools are urgently needed. In this study, we developed a SARS-CoV-2spike (S1) protein enzyme-linked immunosorbent assay (ELISA) to detect SARS-CoV-2-specific antibodies. The SARS-CoV-2 S1 ELISA was found to be specific [97.8% (95% CI, 96.7% - 98.5%)], reproducible and precise (intra-assay coefficient of variability (CV) 5.3%, inter-assay CV 7.9%). A standard curve and the interpolation of arbitrary ELISA units per milliliter served to reduce the variability between different tests and operators. Cross-reactivity to other humancoronaviruses was addressed by using sera positive for MERS-CoV- and hCoV HKU1-specific antibodies. Monitoring antibody development in various samples of twenty-three and single samples of twenty-nine coronavirus disease 2019 (COVID-19) patients revealed seroconversion and neutralizing antibodies against authentic SARS-CoV-2 in all cases. The comparison of the SARS-CoV-2 (S1) ELISA with a commercially available assay showed a better sensitivity for the in-house ELISA. The results demonstrate a high reproducibility, specificity and sensitivity of the newly developed ELISA, which is suitable for the detection of SARS-CoV-2 S1 protein-specific antibody responses.
Authors: Zhanwei Liang; Tao Peng; Xueshima Jiao; Yang Zhao; Jie Xie; You Jiang; Bo Meng; Xiang Fang; Xiaoping Yu; Xinhua Dai Journal: Biosensors (Basel) Date: 2022-02-07
Authors: Marcelo S Conzentino; Ana C A Gonçalves; Nigella M Paula; Fabiane G M Rego; Dalila L Zanette; Mateus N Aoki; Jeanine M Nardin; Luciano Fernandes Huergo Journal: Braz J Microbiol Date: 2022-04-14 Impact factor: 2.214