Validation of a combined ELISA to detect IgG, IgA and IgM antibody responses to SARS-CoV-2 in mild or moderate non-hospitalised patients

AM Cook; SE Faustini; L.J.  Williams; AF Cunningham; MT Drayson; AM Shields; D.  Kay; T.  Plant; A Huissoon; G.  Wallis; S.  Beck; SE Jossi; M.  Perez-Toledo; M.L.  Newby; J.D.  Allen; M.  Crispin; S.  Harding; AG Richter

doi:10.1016/j.jim.2021.113046

Validation of a combined ELISA to detect IgG, IgA and IgM antibody responses to SARS-CoV-2 in mild or moderate non-hospitalised patients

AM Cook, SE Faustini, L.J. Williams, AF Cunningham, MT Drayson, AM Shields, D. Kay, T. Plant, A Huissoon, G. Wallis, S. Beck, SE Jossi, M. Perez-Toledo, M.L. Newby, J.D. Allen, M. Crispin, S. Harding, AG Richter

Research output: Contribution to journal › Article › peer-review

2 Citations (Scopus)

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.

Original language	English
Article number	113046
Number of pages	7
Journal	Journal of Immunological Methods
Volume	494
Early online date	26 Mar 2021
DOIs	https://doi.org/10.1016/j.jim.2021.113046
Publication status	Published - Jul 2021

Bibliographical note

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.

Keywords

COVID-19
Dried blood spot
Low prevalence
Mild disease
Serological
Trimeric spike protein

ASJC Scopus subject areas

Immunology and Allergy
Immunology

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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https://eprints.soton.ac.uk/448852/Licence: Creative Commons: Attribution-NonCommercial-NoDerivs (CC BY-NC-ND)

Cite this

Cook, AM., Faustini, SE., Williams, L. J., Cunningham, AF., Drayson, MT., Shields, AM., Kay, D., Plant, T., Huissoon, A., Wallis, G., Beck, S., Jossi, SE., Perez-Toledo, M., Newby, M. L., Allen, J. D., Crispin, M., Harding, S., & Richter, AG. (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, Article 113046. Advance online publication. https://doi.org/10.1016/j.jim.2021.113046

@article{ba18283247e6401586c712410746c961,

title = "Validation of a combined ELISA to detect IgG, IgA and IgM antibody responses to SARS-CoV-2 in mild or moderate non-hospitalised patients",

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.",

keywords = "COVID-19, Dried blood spot, Low prevalence, Mild disease, Serological, Trimeric spike protein",

author = "AM Cook and SE Faustini and L.J. Williams and AF Cunningham and MT Drayson and AM Shields and D. Kay and T. Plant and A Huissoon and G. Wallis and S. Beck and SE Jossi and M. Perez-Toledo and M.L. Newby and J.D. Allen and M. Crispin and S. Harding and AG Richter",

note = "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.",

year = "2021",

month = jul,

doi = "10.1016/j.jim.2021.113046",

language = "English",

volume = "494",

journal = "Journal of Immunological Methods",

issn = "0022-1759",

publisher = "Elsevier",

}

Cook, AM , Faustini, SE, Williams, LJ, Cunningham, AF , Drayson, MT , Shields, AM, Kay, D, Plant, T, Huissoon, A, Wallis, G, Beck, S, Jossi, SE, Perez-Toledo, M, Newby, ML, Allen, JD, Crispin, M, Harding, S & Richter, AG 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, vol. 494, 113046. https://doi.org/10.1016/j.jim.2021.113046

TY - JOUR

T1 - Validation of a combined ELISA to detect IgG, IgA and IgM antibody responses to SARS-CoV-2 in mild or moderate non-hospitalised patients

AU - Cook, AM

AU - Faustini, SE

AU - Williams, L.J.

AU - Cunningham, AF

AU - Drayson, MT

AU - Shields, AM

AU - Kay, D.

AU - Plant, T.

AU - Huissoon, A

AU - Wallis, G.

AU - Beck, S.

AU - Jossi, SE

AU - Perez-Toledo, M.

AU - Newby, M.L.

AU - Allen, J.D.

AU - Crispin, M.

AU - Harding, S.

AU - Richter, AG

N1 - 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.

PY - 2021/7

Y1 - 2021/7

N2 - 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.

AB - 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.

KW - COVID-19

KW - Dried blood spot

KW - Low prevalence

KW - Mild disease

KW - Serological

KW - Trimeric spike protein

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DO - 10.1016/j.jim.2021.113046

M3 - Article

C2 - 33775672

SN - 0022-1759

VL - 494

JO - Journal of Immunological Methods

JF - Journal of Immunological Methods

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ER -

Validation of a combined ELISA to detect IgG, IgA and IgM antibody responses to SARS-CoV-2 in mild or moderate non-hospitalised patients

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

UN SDGs

Access to Document

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Cite this