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A highly specific and sensitive serological assay detects SARS-CoV-2 antibody levels in COVID-19 patients that correlate with neutralization.
Infection. 2021 Feb; 49(1):75-82.I

Abstract

OBJECTIVE

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic challenges national health systems and the global economy. Monitoring of infection rates and seroprevalence can guide public health measures to combat the pandemic. This depends on reliable tests on active and former infections. Here, we set out to develop and validate a specific and sensitive enzyme linked immunosorbent assay (ELISA) for detection of anti-SARS-CoV-2 antibody levels.

METHODS

In our ELISA, we used SARS-CoV-2 receptor-binding domain (RBD) and a stabilized version of the spike (S) ectodomain as antigens. We assessed sera from patients infected with seasonal coronaviruses, SARS-CoV-2 and controls. We determined and monitored IgM-, IgA- and IgG-antibody responses towards these antigens. In addition, for a panel of 22 sera, virus neutralization and ELISA parameters were measured and correlated.

RESULTS

The RBD-based ELISA detected SARS-CoV-2-directed antibodies, did not cross-react with seasonal coronavirus antibodies and correlated with virus neutralization (R2 = 0.89). Seroconversion started at 5 days after symptom onset and led to robust antibody levels at 10 days after symptom onset. We demonstrate high specificity (99.3%; N = 1000) and sensitivity (92% for IgA, 96% for IgG and 98% for IgM; > 10 days after PCR-proven infection; N = 53) in serum.

CONCLUSIONS

With the described RBD-based ELISA protocol, we provide a reliable test for seroepidemiological surveys. Due to high specificity and strong correlation with virus neutralization, the RBD ELISA holds great potential to become a preferred tool to assess thresholds of protective immunity after infection and vaccination.

Authors+Show Affiliations

Institute for Medical Microbiology and Hygiene, University of Regensburg, Regensburg, Germany. david.peterhoff@ur.de.Institute for Clinical Microbiology and Hygiene, University Hospital Regensburg, Regensburg, Germany.Institute for Clinical Microbiology and Hygiene, University Hospital Regensburg, Regensburg, Germany.Institute for Medical Microbiology and Hygiene, University of Regensburg, Regensburg, Germany.Institute for Medical Microbiology and Hygiene, University of Regensburg, Regensburg, Germany.Institute for Clinical Microbiology and Hygiene, University Hospital Regensburg, Regensburg, Germany.Institute for Clinical Microbiology and Hygiene, University Hospital Regensburg, Regensburg, Germany.Institute for Clinical Microbiology and Hygiene, University Hospital Regensburg, Regensburg, Germany.Institute for Clinical Microbiology and Hygiene, University Hospital Regensburg, Regensburg, Germany.Institute for Clinical Microbiology and Hygiene, University Hospital Regensburg, Regensburg, Germany.Institute for Medical Microbiology and Hygiene, University of Regensburg, Regensburg, Germany.Institute for Clinical Microbiology and Hygiene, University Hospital Regensburg, Regensburg, Germany.Institute for Clinical Microbiology and Hygiene, University Hospital Regensburg, Regensburg, Germany.Institute for Clinical Microbiology and Hygiene, University Hospital Regensburg, Regensburg, Germany.Department for Infection Control and Infectious Diseases, University Hospital Regensburg, Regensburg, Germany. Emergency Department, University Hospital Regensburg, Regensburg, Germany.Department of Internal Medicine II, University Hospital Regensburg, Regensburg, Germany.Department of Internal Medicine II, University Hospital Regensburg, Regensburg, Germany.Department of Anesthesiology, University Hospital Regensburg, Regensburg, Germany.Department for Infection Control and Infectious Diseases, University Hospital Regensburg, Regensburg, Germany.Praxiszentrum Alte Mälzerei, Regensburg, Germany.Institute of Clinical Chemistry and Laboratory Medicine, Transfusion Medicine, University Hospital Regensburg, Regensburg, Germany.Institute of Clinical Chemistry and Laboratory Medicine, Transfusion Medicine, University Hospital Regensburg, Regensburg, Germany.Department of Internal Medicine I, University Hospital Regensburg, Regensburg, Germany.Department of Internal Medicine I, University Hospital Regensburg, Regensburg, Germany.Institute of Clinical Chemistry and Laboratory Medicine, University Hospital Regensburg, Regensburg, Germany.Kreisklinik Trostberg, Trostberg, Germany.Center for Clinical Studies, University Hospital Regensburg, Regensburg, Germany.Institute for Medical Microbiology and Hygiene, University of Regensburg, Regensburg, Germany.Department of Anesthesiology, University Hospital Regensburg, Regensburg, Germany.Department for Infection Control and Infectious Diseases, University Hospital Regensburg, Regensburg, Germany.Institute for Clinical Microbiology and Hygiene, University Hospital Regensburg, Regensburg, Germany.Institute for Medical Microbiology and Hygiene, University of Regensburg, Regensburg, Germany. Institute for Clinical Microbiology and Hygiene, University Hospital Regensburg, Regensburg, Germany.Institute for Medical Microbiology and Hygiene, University of Regensburg, Regensburg, Germany. Institute for Clinical Microbiology and Hygiene, University Hospital Regensburg, Regensburg, Germany.Institute for Medical Microbiology and Hygiene, University of Regensburg, Regensburg, Germany. barbara.schmidt@ukr.de. Institute for Clinical Microbiology and Hygiene, University Hospital Regensburg, Regensburg, Germany. barbara.schmidt@ukr.de.Institute for Medical Microbiology and Hygiene, University of Regensburg, Regensburg, Germany. ralf.wagner@ur.de. Institute for Clinical Microbiology and Hygiene, University Hospital Regensburg, Regensburg, Germany. ralf.wagner@ur.de.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

32827125

Citation

Peterhoff, David, et al. "A Highly Specific and Sensitive Serological Assay Detects SARS-CoV-2 Antibody Levels in COVID-19 Patients That Correlate With Neutralization." Infection, vol. 49, no. 1, 2021, pp. 75-82.
Peterhoff D, Glück V, Vogel M, et al. A highly specific and sensitive serological assay detects SARS-CoV-2 antibody levels in COVID-19 patients that correlate with neutralization. Infection. 2021;49(1):75-82.
Peterhoff, D., Glück, V., Vogel, M., Schuster, P., Schütz, A., Neubert, P., Albert, V., Frisch, S., Kiessling, M., Pervan, P., Werner, M., Ritter, N., Babl, L., Deichner, M., Hanses, F., Lubnow, M., Müller, T., Lunz, D., Hitzenbichler, F., ... Wagner, R. (2021). A highly specific and sensitive serological assay detects SARS-CoV-2 antibody levels in COVID-19 patients that correlate with neutralization. Infection, 49(1), 75-82. https://doi.org/10.1007/s15010-020-01503-7
Peterhoff D, et al. A Highly Specific and Sensitive Serological Assay Detects SARS-CoV-2 Antibody Levels in COVID-19 Patients That Correlate With Neutralization. Infection. 2021;49(1):75-82. PubMed PMID: 32827125.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - A highly specific and sensitive serological assay detects SARS-CoV-2 antibody levels in COVID-19 patients that correlate with neutralization. AU - Peterhoff,David, AU - Glück,Vivian, AU - Vogel,Matthias, AU - Schuster,Philipp, AU - Schütz,Anja, AU - Neubert,Philip, AU - Albert,Veruschka, AU - Frisch,Stefanie, AU - Kiessling,Mara, AU - Pervan,Philip, AU - Werner,Maren, AU - Ritter,Nicole, AU - Babl,Leon, AU - Deichner,Maria, AU - Hanses,Frank, AU - Lubnow,Matthias, AU - Müller,Thomas, AU - Lunz,Dirk, AU - Hitzenbichler,Florian, AU - Audebert,Franz, AU - Hähnel,Viola, AU - Offner,Robert, AU - Müller,Martina, AU - Schmid,Stephan, AU - Burkhardt,Ralph, AU - Glück,Thomas, AU - Koller,Michael, AU - Niller,Hans Helmut, AU - Graf,Bernhard, AU - Salzberger,Bernd, AU - Wenzel,Jürgen J, AU - Jantsch,Jonathan, AU - Gessner,André, AU - Schmidt,Barbara, AU - Wagner,Ralf, Y1 - 2020/08/21/ PY - 2020/07/01/received PY - 2020/08/07/accepted PY - 2020/8/23/pubmed PY - 2021/2/10/medline PY - 2020/8/23/entrez KW - Antibody test KW - Assay validation KW - COVID-19 KW - ELISA KW - Receptor binding domain KW - S protein KW - SARS-CoV-2 KW - Serology KW - Spike protein KW - Virus neutralization SP - 75 EP - 82 JF - Infection JO - Infection VL - 49 IS - 1 N2 - OBJECTIVE: The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic challenges national health systems and the global economy. Monitoring of infection rates and seroprevalence can guide public health measures to combat the pandemic. This depends on reliable tests on active and former infections. Here, we set out to develop and validate a specific and sensitive enzyme linked immunosorbent assay (ELISA) for detection of anti-SARS-CoV-2 antibody levels. METHODS: In our ELISA, we used SARS-CoV-2 receptor-binding domain (RBD) and a stabilized version of the spike (S) ectodomain as antigens. We assessed sera from patients infected with seasonal coronaviruses, SARS-CoV-2 and controls. We determined and monitored IgM-, IgA- and IgG-antibody responses towards these antigens. In addition, for a panel of 22 sera, virus neutralization and ELISA parameters were measured and correlated. RESULTS: The RBD-based ELISA detected SARS-CoV-2-directed antibodies, did not cross-react with seasonal coronavirus antibodies and correlated with virus neutralization (R2 = 0.89). Seroconversion started at 5 days after symptom onset and led to robust antibody levels at 10 days after symptom onset. We demonstrate high specificity (99.3%; N = 1000) and sensitivity (92% for IgA, 96% for IgG and 98% for IgM; > 10 days after PCR-proven infection; N = 53) in serum. CONCLUSIONS: With the described RBD-based ELISA protocol, we provide a reliable test for seroepidemiological surveys. Due to high specificity and strong correlation with virus neutralization, the RBD ELISA holds great potential to become a preferred tool to assess thresholds of protective immunity after infection and vaccination. SN - 1439-0973 UR - https://www.unboundmedicine.com/medline/citation/32827125/A_highly_specific_and_sensitive_serological_assay_detects_SARS_CoV_2_antibody_levels_in_COVID_19_patients_that_correlate_with_neutralization_ L2 - https://doi.org/10.1007/s15010-020-01503-7 DB - PRIME DP - Unbound Medicine ER -