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Evaluating SARS-CoV-2 spike and nucleocapsid proteins as targets for antibody detection in severe and mild COVID-19 cases using a Luminex bead-based assay.
J Virol Methods. 2021 02; 288:114025.JV

Abstract

Large-scale serosurveillance of severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2) will only be possible if serological tests are sufficiently reliable, rapid and affordable. Many assays are either labour-intensive and require specialised facilities (e.g. virus neutralization assays), or are expensive with suboptimal specificity (e.g. commercial ELISAs and RDTs). Bead-based assays offer a cost-effective alternative and allow for multiplexing to test for antibodies against multiple antigens and against other pathogens. Here, we compare the performance of spike (S) and nucleocapsid (NP) antigens for the detection of SARS-CoV-2 specific IgG, IgM and IgA antibodies in a panel of sera that includes recent (up to six weeks after symptom onset, severe n = 44; and mild cases n = 52) and old infections (five months after symptom onset, mild n = 104), using a Luminex-bead based assay and comparison to a virus neutralization test. While we show that neutralizing antibody levels are significantly lower in mild than in severe cases, we demonstrate that a combination of the recombinant nucleocapsid protein (NP) and receptor-binding domain (RBD) results in highly specific (99 %) IgG antibody detection five months after infection in 96 % of cases. Although most severe Covid-19 cases developed a clear IgM and IgA response, titers fell below the detection threshold in more than 20 % of mild cases in our bead-based assay. In conclusion, our data supports the use of RBD and NP for the development of SARS-CoV-2 serological IgG bead-based assays.

Authors+Show Affiliations

Outbreak Research Team, Institute of Tropical Medicine, Antwerp, Belgium. Electronic address: joachim_marien@hotmail.com.Virology Unit, Department of Biomedical Sciences, Institute of Tropical Medicine, Antwerp, Belgium.Virology Unit, Department of Biomedical Sciences, Institute of Tropical Medicine, Antwerp, Belgium.Virology Unit, Department of Biomedical Sciences, Institute of Tropical Medicine, Antwerp, Belgium.Virology Unit, Department of Biomedical Sciences, Institute of Tropical Medicine, Antwerp, Belgium.Department of Clinical Sciences, Institute of Tropical Medicine, Antwerp, Belgium.Outbreak Research Team, Institute of Tropical Medicine, Antwerp, Belgium.Epidemiology and Public Health, Sciensano, Brussels, Belgium.Epidemiology and Public Health, Sciensano, Brussels, Belgium.Immune Response, Sciensano, Brussels, Belgium.University Hospital Antwerp, Antwerp, Belgium.Department of Clinical Sciences, Institute of Tropical Medicine, Antwerp, Belgium.Virology Unit, Department of Biomedical Sciences, Institute of Tropical Medicine, Antwerp, Belgium; University of Antwerp, Antwerp, Belgium. Electronic address: KArien@itg.be.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

33227340

Citation

Mariën, Joachim, et al. "Evaluating SARS-CoV-2 Spike and Nucleocapsid Proteins as Targets for Antibody Detection in Severe and Mild COVID-19 Cases Using a Luminex Bead-based Assay." Journal of Virological Methods, vol. 288, 2021, p. 114025.
Mariën J, Ceulemans A, Michiels J, et al. Evaluating SARS-CoV-2 spike and nucleocapsid proteins as targets for antibody detection in severe and mild COVID-19 cases using a Luminex bead-based assay. J Virol Methods. 2021;288:114025.
Mariën, J., Ceulemans, A., Michiels, J., Heyndrickx, L., Kerkhof, K., Foque, N., Widdowson, M. A., Mortgat, L., Duysburgh, E., Desombere, I., Jansens, H., Van Esbroeck, M., & Ariën, K. K. (2021). Evaluating SARS-CoV-2 spike and nucleocapsid proteins as targets for antibody detection in severe and mild COVID-19 cases using a Luminex bead-based assay. Journal of Virological Methods, 288, 114025. https://doi.org/10.1016/j.jviromet.2020.114025
Mariën J, et al. Evaluating SARS-CoV-2 Spike and Nucleocapsid Proteins as Targets for Antibody Detection in Severe and Mild COVID-19 Cases Using a Luminex Bead-based Assay. J Virol Methods. 2021;288:114025. PubMed PMID: 33227340.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Evaluating SARS-CoV-2 spike and nucleocapsid proteins as targets for antibody detection in severe and mild COVID-19 cases using a Luminex bead-based assay. AU - Mariën,Joachim, AU - Ceulemans,Ann, AU - Michiels,Johan, AU - Heyndrickx,Leo, AU - Kerkhof,Karen, AU - Foque,Nikki, AU - Widdowson,Marc-Alain, AU - Mortgat,Laure, AU - Duysburgh,Els, AU - Desombere,Isabelle, AU - Jansens,Hilde, AU - Van Esbroeck,Marjan, AU - Ariën,Kevin K, Y1 - 2020/11/20/ PY - 2020/08/04/received PY - 2020/11/16/revised PY - 2020/11/18/accepted PY - 2020/11/24/pubmed PY - 2020/11/24/medline PY - 2020/11/23/entrez KW - Bead-based assay KW - Luminex antibody test KW - Sars-CoV-2 KW - Serosurveillance KW - Virus neutralization test SP - 114025 EP - 114025 JF - Journal of virological methods JO - J Virol Methods VL - 288 N2 - Large-scale serosurveillance of severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2) will only be possible if serological tests are sufficiently reliable, rapid and affordable. Many assays are either labour-intensive and require specialised facilities (e.g. virus neutralization assays), or are expensive with suboptimal specificity (e.g. commercial ELISAs and RDTs). Bead-based assays offer a cost-effective alternative and allow for multiplexing to test for antibodies against multiple antigens and against other pathogens. Here, we compare the performance of spike (S) and nucleocapsid (NP) antigens for the detection of SARS-CoV-2 specific IgG, IgM and IgA antibodies in a panel of sera that includes recent (up to six weeks after symptom onset, severe n = 44; and mild cases n = 52) and old infections (five months after symptom onset, mild n = 104), using a Luminex-bead based assay and comparison to a virus neutralization test. While we show that neutralizing antibody levels are significantly lower in mild than in severe cases, we demonstrate that a combination of the recombinant nucleocapsid protein (NP) and receptor-binding domain (RBD) results in highly specific (99 %) IgG antibody detection five months after infection in 96 % of cases. Although most severe Covid-19 cases developed a clear IgM and IgA response, titers fell below the detection threshold in more than 20 % of mild cases in our bead-based assay. In conclusion, our data supports the use of RBD and NP for the development of SARS-CoV-2 serological IgG bead-based assays. SN - 1879-0984 UR - https://www.unboundmedicine.com/medline/citation/33227340/Evaluating_SARS_CoV_2_spike_and_nucleocapsid_proteins_as_targets_for_antibody_detection_in_severe_and_mild_COVID_19_cases_using_a_Luminex_bead_based_assay_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0166-0934(20)30277-9 DB - PRIME DP - Unbound Medicine ER -