Validation of a combined ELISA to detect IgG, IgA and IgM antibody responses to SARS-CoV-2 in mild or moderate non-hospitalised patients
Validation of a combined ELISA to detect IgG, IgA and IgM antibody responses to SARS-CoV-2 in mild or moderate non-hospitalised patients
Background: Frequently SARS-CoV-2 results in mild or moderate disease with potentially lower concentrations of antibodies compared to those that are hospitalised. Here, we validated an ELISA using SARS-CoV-2 trimeric spike glycoprotein, with targeted detection of IgG, IgA and IgM (IgGAM) using serum and dried blood spots (DBS) from adults with mild or moderate disease. Methods: Targeting the SARS-CoV-2 trimeric spike, a combined anti-IgG, IgA and IgM serology ELISA assay was developed using 62 PCR-confirmed non-hospitalised, mild or moderate COVID-19 samples, ≥14 days post symptom onset and 624 COVID-19 negative samples. The assay was validated using 73 PCR-confirmed non-hospitalised, mild or moderate COVID-19 samples, ≥14 days post symptom onset and 359 COVID-19 negative serum samples with an additional 81 DBSs. The assay was further validated in 226 PCR-confirmed non-hospitalised, mild or moderate COVID-19 samples, ≥14 days post symptom onset and 426 COVID-19 negative clinical samples. Results: A sensitivity and specificity of 98.6% (95% CI, 92.6–100.0), 98.3% (95% CI, 96.4–99.4), respectively, was observed following validation of the SARS-CoV-2 ELISA. No cross-reactivities with endemic coronaviruses or other human viruses were observed, and no change in results were recorded for interfering substances. The assay was stable at temperature extremes and components were stable for 15 days once opened. A matrix comparison showed DBS to correlate with serum results. Clinical validation of the assay reported a sensitivity of 94.7% (95% CI, 90.9–97.2%) and a specificity of 98.4% (95% CI, 96.6–99.3%). Conclusions: The human anti-IgGAM SARS-CoV-2 ELISA provides accurate and sensitive detection of SARS-CoV-2 antibodies in non-hospitalised adults with mild or moderate disease. The use of dried blood spots makes the assay accessible to the wider community.
COVID-19, Dried blood spot, Low prevalence, Mild disease, Serological, Trimeric spike protein
1-7
Cook, A. M.
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Faustini, S. E.
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Williams, L. J.
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Cunningham, A. F.
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Drayson, M. T.
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Shields, A. M.
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Kay, D.
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Taylor, L.
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Plant, T.
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Huissoon, A.
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Wallis, G.
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Beck, S.
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Jossi, S. E.
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Perez-Toledo, M.
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Newby, M. L.
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Allen, J. D.
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Crispin, M.
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Harding, S.
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Richter, A. G.
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1 July 2021
Cook, A. M.
40971c28-32c9-4294-9447-c592024434ec
Faustini, S. E.
fc25e886-bb2c-4c9c-a421-df8e00e6b1ba
Williams, L. J.
40fbbf1f-4147-4e57-a0ca-c46dc8038af8
Cunningham, A. F.
46ea6603-b3e9-492e-92e2-0018103566fc
Drayson, M. T.
24a45661-4fb4-4063-b918-c519ac4f06ec
Shields, A. M.
0ea27afe-3e23-472d-b8d8-fc0ed49e93dd
Kay, D.
7c403f95-8d6a-4b19-90ab-ea9f1a2cf35b
Taylor, L.
23d42821-0be9-43fa-8793-af51dbfbcea5
Plant, T.
70eb86b0-3d61-47cf-b39c-66835fc4c7d9
Huissoon, A.
9fa377fb-d33d-41a3-8b30-a14c7f3b8768
Wallis, G.
89b55046-29ce-4131-8c85-0ab5ed0a722f
Beck, S.
67ec6ea3-2aed-415b-894b-55eefb959479
Jossi, S. E.
e2b4dd30-7a37-4243-8bbf-9132cc7b41fd
Perez-Toledo, M.
db9dfcc0-d3a0-4c9d-9606-f2370574e3d3
Newby, M. L.
417cba47-6a6f-42b9-8b9c-640f0518c621
Allen, J. D.
c89d5569-7659-4835-b535-c9586e956b3a
Crispin, M.
cd980957-0943-4b89-b2b2-710f01f33bc9
Harding, S.
b874100d-5b0e-4d37-b507-96d6c622fb88
Richter, A. G.
a8d5bc53-c51b-463d-af6b-ca8b8a5412d0
Cook, A. M., Faustini, S. E., Williams, L. J., Cunningham, A. F., Drayson, M. T., Shields, A. M., Kay, D., Taylor, L., Plant, T., Huissoon, A., Wallis, G., Beck, S., Jossi, S. E., Perez-Toledo, M., Newby, M. L., Allen, J. D., Crispin, M., Harding, S. and Richter, A. G.
(2021)
Validation of a combined ELISA to detect IgG, IgA and IgM antibody responses to SARS-CoV-2 in mild or moderate non-hospitalised patients.
Journal of Immunological Methods, 494, , [113046].
(doi:10.1016/j.jim.2021.113046).
Abstract
Background: Frequently SARS-CoV-2 results in mild or moderate disease with potentially lower concentrations of antibodies compared to those that are hospitalised. Here, we validated an ELISA using SARS-CoV-2 trimeric spike glycoprotein, with targeted detection of IgG, IgA and IgM (IgGAM) using serum and dried blood spots (DBS) from adults with mild or moderate disease. Methods: Targeting the SARS-CoV-2 trimeric spike, a combined anti-IgG, IgA and IgM serology ELISA assay was developed using 62 PCR-confirmed non-hospitalised, mild or moderate COVID-19 samples, ≥14 days post symptom onset and 624 COVID-19 negative samples. The assay was validated using 73 PCR-confirmed non-hospitalised, mild or moderate COVID-19 samples, ≥14 days post symptom onset and 359 COVID-19 negative serum samples with an additional 81 DBSs. The assay was further validated in 226 PCR-confirmed non-hospitalised, mild or moderate COVID-19 samples, ≥14 days post symptom onset and 426 COVID-19 negative clinical samples. Results: A sensitivity and specificity of 98.6% (95% CI, 92.6–100.0), 98.3% (95% CI, 96.4–99.4), respectively, was observed following validation of the SARS-CoV-2 ELISA. No cross-reactivities with endemic coronaviruses or other human viruses were observed, and no change in results were recorded for interfering substances. The assay was stable at temperature extremes and components were stable for 15 days once opened. A matrix comparison showed DBS to correlate with serum results. Clinical validation of the assay reported a sensitivity of 94.7% (95% CI, 90.9–97.2%) and a specificity of 98.4% (95% CI, 96.6–99.3%). Conclusions: The human anti-IgGAM SARS-CoV-2 ELISA provides accurate and sensitive detection of SARS-CoV-2 antibodies in non-hospitalised adults with mild or moderate disease. The use of dried blood spots makes the assay accessible to the wider community.
Text
validation of a combined ELISA to detect IgG, IgA and IgM antibody responses to SARS-cOv-2
- Accepted Manuscript
More information
Accepted/In Press date: 20 March 2021
e-pub ahead of print date: 26 March 2021
Published date: 1 July 2021
Additional Information:
Funding Information:
We thank Lia van der Hoek for seasonal coronaviruses samples used in the cross reactivity study from the Amsterdam Cohort Studies (ACS) on HIV infection and AIDS. The ACS is a collaboration among the Public Health Service of Amsterdam, the Amsterdam University Medical Center of the University of Amsterdam, the Sanquin Blood Supply Foundation, the Medical Center Jan van Goyen and the HIV Focus Center of the DC-Clinics. It is part of the Netherlands HIV Monitoring Foundation and financially supported by the Center for Infectious Disease Control of the Netherlands National Institute for Public Health and the Environment. The authors thank all ACS participants for their contribution, as well as the ACS study nurses, data managers and lab technicians.
Funding Information:
We thank Lia van der Hoek for seasonal coronaviruses samples used in the cross reactivity study from the Amsterdam Cohort Studies (ACS) on HIV infection and AIDS. The ACS is a collaboration among the Public Health Service of Amsterdam, the Amsterdam University Medical Center of the University of Amsterdam, the Sanquin Blood Supply Foundation, the Medical Center Jan van Goyen and the HIV Focus Center of the DC-Clinics. It is part of the Netherlands HIV Monitoring Foundation and financially supported by the Center for Infectious Disease Control of the Netherlands National Institute for Public Health and the Environment . The authors thank all ACS participants for their contribution, as well as the ACS study nurses, data managers and lab technicians.
Publisher Copyright:
© 2021
Copyright:
Copyright 2021 Elsevier B.V., All rights reserved.
Keywords:
COVID-19, Dried blood spot, Low prevalence, Mild disease, Serological, Trimeric spike protein
Identifiers
Local EPrints ID: 448852
URI: http://eprints.soton.ac.uk/id/eprint/448852
ISSN: 0022-1759
PURE UUID: a1376ea8-9532-43d9-b929-f522248bb3d4
Catalogue record
Date deposited: 06 May 2021 16:34
Last modified: 18 Mar 2024 05:27
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Contributors
Author:
A. M. Cook
Author:
S. E. Faustini
Author:
L. J. Williams
Author:
A. F. Cunningham
Author:
M. T. Drayson
Author:
A. M. Shields
Author:
D. Kay
Author:
L. Taylor
Author:
T. Plant
Author:
A. Huissoon
Author:
G. Wallis
Author:
S. Beck
Author:
S. E. Jossi
Author:
M. Perez-Toledo
Author:
M. L. Newby
Author:
S. Harding
Author:
A. G. Richter
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