tetano
Editor, Senior Moderator
J Immunol Methods
. 2021 Jan 4;112958.
doi: 10.1016/j.jim.2021.112958. Online ahead of print.
Development and characterization of an indirect ELISA to detect SARS-CoV - 2 spike protein-specific antibodies
Verena Kr?hling[SUP] 1 [/SUP], Sandro Halwe[SUP] 2 [/SUP], Cornelius Rohde[SUP] 2 [/SUP], Dirk Becker[SUP] 2 [/SUP], Susanne Bergh?fer[SUP] 3 [/SUP], Christine Dahlke[SUP] 4 [/SUP], Markus Eickmann[SUP] 3 [/SUP], Meryem S Ercanoglu[SUP] 5 [/SUP], Lutz Gieselmann[SUP] 5 [/SUP], Astrid Herwig[SUP] 3 [/SUP], Alexandra Kupke[SUP] 2 [/SUP], Helena M?ller[SUP] 3 [/SUP], Petra Neubauer-R?del[SUP] 3 [/SUP], Florian Klein[SUP] 6 [/SUP], Christian Keller[SUP] 7 [/SUP], Stephan Becker[SUP] 2 [/SUP]
Affiliations
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.
Keywords: COVID-19; Clinical diagnostic; ELISA; SARS-CoV-2; Spike protein.
. 2021 Jan 4;112958.
doi: 10.1016/j.jim.2021.112958. Online ahead of print.
Development and characterization of an indirect ELISA to detect SARS-CoV - 2 spike protein-specific antibodies
Verena Kr?hling[SUP] 1 [/SUP], Sandro Halwe[SUP] 2 [/SUP], Cornelius Rohde[SUP] 2 [/SUP], Dirk Becker[SUP] 2 [/SUP], Susanne Bergh?fer[SUP] 3 [/SUP], Christine Dahlke[SUP] 4 [/SUP], Markus Eickmann[SUP] 3 [/SUP], Meryem S Ercanoglu[SUP] 5 [/SUP], Lutz Gieselmann[SUP] 5 [/SUP], Astrid Herwig[SUP] 3 [/SUP], Alexandra Kupke[SUP] 2 [/SUP], Helena M?ller[SUP] 3 [/SUP], Petra Neubauer-R?del[SUP] 3 [/SUP], Florian Klein[SUP] 6 [/SUP], Christian Keller[SUP] 7 [/SUP], Stephan Becker[SUP] 2 [/SUP]
Affiliations
- PMID: 33412174
- DOI: 10.1016/j.jim.2021.112958
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.
Keywords: COVID-19; Clinical diagnostic; ELISA; SARS-CoV-2; Spike protein.