Systematic evaluation of SARS-CoV-2 antigens enables a highly specific and sensitive multiplex serological COVID-19 assay
(2021) In Clinical and Translational Immunology 10(7).- Abstract
Objective: The COVID-19 pandemic poses an immense need for accurate, sensitive and high-throughput clinical tests, and serological assays are needed for both overarching epidemiological studies and evaluating vaccines. Here, we present the development and validation of a high-throughput multiplex bead-based serological assay. Methods: More than 100 representations of SARS-CoV-2 proteins were included for initial evaluation, including antigens produced in bacterial and mammalian hosts as well as synthetic peptides. The five best-performing antigens, three representing the spike glycoprotein and two representing the nucleocapsid protein, were further evaluated for detection of IgG antibodies in samples from 331 COVID-19 patients and... (More)
Objective: The COVID-19 pandemic poses an immense need for accurate, sensitive and high-throughput clinical tests, and serological assays are needed for both overarching epidemiological studies and evaluating vaccines. Here, we present the development and validation of a high-throughput multiplex bead-based serological assay. Methods: More than 100 representations of SARS-CoV-2 proteins were included for initial evaluation, including antigens produced in bacterial and mammalian hosts as well as synthetic peptides. The five best-performing antigens, three representing the spike glycoprotein and two representing the nucleocapsid protein, were further evaluated for detection of IgG antibodies in samples from 331 COVID-19 patients and convalescents, and in 2090 negative controls sampled before 2020. Results: Three antigens were finally selected, represented by a soluble trimeric form and the S1-domain of the spike glycoprotein as well as by the C-terminal domain of the nucleocapsid. The sensitivity for these three antigens individually was found to be 99.7%, 99.1% and 99.7%, and the specificity was found to be 98.1%, 98.7% and 95.7%. The best assay performance was although achieved when utilising two antigens in combination, enabling a sensitivity of up to 99.7% combined with a specificity of 100%. Requiring any two of the three antigens resulted in a sensitivity of 99.7% and a specificity of 99.4%. Conclusion: These observations demonstrate that a serological test based on a combination of several SARS-CoV-2 antigens enables a highly specific and sensitive multiplex serological COVID-19 assay.
(Less)
- author
- organization
- publishing date
- 2021
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- COVID-19, IgG, multiplex, SARS-CoV-2, serological assay
- in
- Clinical and Translational Immunology
- volume
- 10
- issue
- 7
- article number
- e1312
- publisher
- John Wiley & Sons Inc.
- external identifiers
-
- scopus:85111256975
- pmid:34295471
- ISSN
- 2050-0068
- DOI
- 10.1002/cti2.1312
- language
- English
- LU publication?
- yes
- id
- 73f69d92-0a20-4fb9-b33d-2c17fa5f0685
- date added to LUP
- 2021-09-01 14:19:02
- date last changed
- 2024-05-04 11:28:29
@article{73f69d92-0a20-4fb9-b33d-2c17fa5f0685,
abstract = {{<p>Objective: The COVID-19 pandemic poses an immense need for accurate, sensitive and high-throughput clinical tests, and serological assays are needed for both overarching epidemiological studies and evaluating vaccines. Here, we present the development and validation of a high-throughput multiplex bead-based serological assay. Methods: More than 100 representations of SARS-CoV-2 proteins were included for initial evaluation, including antigens produced in bacterial and mammalian hosts as well as synthetic peptides. The five best-performing antigens, three representing the spike glycoprotein and two representing the nucleocapsid protein, were further evaluated for detection of IgG antibodies in samples from 331 COVID-19 patients and convalescents, and in 2090 negative controls sampled before 2020. Results: Three antigens were finally selected, represented by a soluble trimeric form and the S1-domain of the spike glycoprotein as well as by the C-terminal domain of the nucleocapsid. The sensitivity for these three antigens individually was found to be 99.7%, 99.1% and 99.7%, and the specificity was found to be 98.1%, 98.7% and 95.7%. The best assay performance was although achieved when utilising two antigens in combination, enabling a sensitivity of up to 99.7% combined with a specificity of 100%. Requiring any two of the three antigens resulted in a sensitivity of 99.7% and a specificity of 99.4%. Conclusion: These observations demonstrate that a serological test based on a combination of several SARS-CoV-2 antigens enables a highly specific and sensitive multiplex serological COVID-19 assay.</p>}},
author = {{Hober, Sophia and Hellström, Cecilia and Olofsson, Jennie and Andersson, Eni and Bergström, Sofia and Jernbom Falk, August and Bayati, Shaghayegh and Mravinacova, Sara and Sjöberg, Ronald and Yousef, Jamil and Skoglund, Lovisa and Kanje, Sara and Berling, Anna and Svensson, Anne Sophie and Jensen, Gabriella and Enstedt, Henric and Afshari, Delaram and Xu, Lan Lan and Zwahlen, Martin and von Feilitzen, Kalle and Hanke, Leo and Murrell, Ben and McInerney, Gerald and Karlsson Hedestam, Gunilla B. and Lendel, Christofer and Roth, Robert G. and Skoog, Ingmar and Svenungsson, Elisabet and Olsson, Tomas and Fogdell-Hahn, Anna and Lindroth, Ylva and Lundgren, Maria and Maleki, Kimia T. and Lagerqvist, Nina and Klingström, Jonas and Da Silva Rodrigues, Rui and Muschiol, Sandra and Bogdanovic, Gordana and Arroyo Mühr, Laila Sara and Eklund, Carina and Lagheden, Camilla and Dillner, Joakim and Sivertsson, Åsa and Havervall, Sebastian and Thålin, Charlotte and Tegel, Hanna and Pin, Elisa and Månberg, Anna and Hedhammar, My and Nilsson, Peter}},
issn = {{2050-0068}},
keywords = {{COVID-19; IgG; multiplex; SARS-CoV-2; serological assay}},
language = {{eng}},
number = {{7}},
publisher = {{John Wiley & Sons Inc.}},
series = {{Clinical and Translational Immunology}},
title = {{Systematic evaluation of SARS-CoV-2 antigens enables a highly specific and sensitive multiplex serological COVID-19 assay}},
url = {{http://dx.doi.org/10.1002/cti2.1312}},
doi = {{10.1002/cti2.1312}},
volume = {{10}},
year = {{2021}},
}