Development of an Enzyme-Linked Immunosorbent Assay (ELISA) for Accurate and Prompt Coronavirus Disease 2019 (COVID-19) Diagnosis Using the Rational Selection of Serological Biomarkers

Prompt COVID-19 diagnosis is urgently required to support infection control measures. Currently available serological tests for measuring SARS-CoV-2 antibodies use different target antigens, although their sensitivity and specificity presents a challenge. We aimed to develop an “in-house” serologica...

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Autores principales:	Theano Lagousi, John Routsias, Vana Spoulou
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	SARS-CoV-2 antibodies serological tests Medicine (General) R5-920
Acceso en línea:	https://doaj.org/article/f029449ab9494f3884ccbea4c8e15902
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spelling	oai:doaj.org-article:f029449ab9494f3884ccbea4c8e159022021-11-25T17:20:21ZDevelopment of an Enzyme-Linked Immunosorbent Assay (ELISA) for Accurate and Prompt Coronavirus Disease 2019 (COVID-19) Diagnosis Using the Rational Selection of Serological Biomarkers10.3390/diagnostics111119702075-4418https://doaj.org/article/f029449ab9494f3884ccbea4c8e159022021-10-01T00:00:00Zhttps://www.mdpi.com/2075-4418/11/11/1970https://doaj.org/toc/2075-4418Prompt COVID-19 diagnosis is urgently required to support infection control measures. Currently available serological tests for measuring SARS-CoV-2 antibodies use different target antigens, although their sensitivity and specificity presents a challenge. We aimed to develop an “in-house” serological ELISA to measure antibodies against SARS-CoV-2 by combining different protein antigens. Sera (<i>n</i> = 44) from COVID-19-confirmed patients were evaluated against different SARS-CoV-2 protein antigens and all potential combinations using ELISA. Patients’ sera were also evaluated against commercially available ELISA diagnostic kits. The mixture containing RBD 2.5 μg/mL, S2 1 μg/mL and N 1.5 μg/mL was found to be the most potent. Plates were incubated with patients’ sera (1:100), and goat anti-human alkaline phosphatase-conjugated IgG, ΙgM and IgA antibody was added. The cut-off value for each assay was determined using the mean optical density plus two standard deviations of pre-pandemic controls. The “in-house” ELISA displayed 91% sensitivity and 97% specificity for IgG antibodies, whereas its sensitivity and specificity for IgM and IgA were 75% and 95% and 73% and 91%, respectively. The “in-house” ELISA developed here combined three SARS-CoV-2 antigens (RBD, S2 and N) as capture antigens and displayed comparable and even higher sensitivity and specificity than otherwise quite reliable commercially available ELISA diagnostic kits.Theano LagousiJohn RoutsiasVana SpoulouMDPI AGarticleSARS-CoV-2antibodiesserological testsMedicine (General)R5-920ENDiagnostics, Vol 11, Iss 1970, p 1970 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	SARS-CoV-2 antibodies serological tests Medicine (General) R5-920
spellingShingle	SARS-CoV-2 antibodies serological tests Medicine (General) R5-920 Theano Lagousi John Routsias Vana Spoulou Development of an Enzyme-Linked Immunosorbent Assay (ELISA) for Accurate and Prompt Coronavirus Disease 2019 (COVID-19) Diagnosis Using the Rational Selection of Serological Biomarkers
description	Prompt COVID-19 diagnosis is urgently required to support infection control measures. Currently available serological tests for measuring SARS-CoV-2 antibodies use different target antigens, although their sensitivity and specificity presents a challenge. We aimed to develop an “in-house” serological ELISA to measure antibodies against SARS-CoV-2 by combining different protein antigens. Sera (<i>n</i> = 44) from COVID-19-confirmed patients were evaluated against different SARS-CoV-2 protein antigens and all potential combinations using ELISA. Patients’ sera were also evaluated against commercially available ELISA diagnostic kits. The mixture containing RBD 2.5 μg/mL, S2 1 μg/mL and N 1.5 μg/mL was found to be the most potent. Plates were incubated with patients’ sera (1:100), and goat anti-human alkaline phosphatase-conjugated IgG, ΙgM and IgA antibody was added. The cut-off value for each assay was determined using the mean optical density plus two standard deviations of pre-pandemic controls. The “in-house” ELISA displayed 91% sensitivity and 97% specificity for IgG antibodies, whereas its sensitivity and specificity for IgM and IgA were 75% and 95% and 73% and 91%, respectively. The “in-house” ELISA developed here combined three SARS-CoV-2 antigens (RBD, S2 and N) as capture antigens and displayed comparable and even higher sensitivity and specificity than otherwise quite reliable commercially available ELISA diagnostic kits.
format	article
author	Theano Lagousi John Routsias Vana Spoulou
author_facet	Theano Lagousi John Routsias Vana Spoulou
author_sort	Theano Lagousi
title	Development of an Enzyme-Linked Immunosorbent Assay (ELISA) for Accurate and Prompt Coronavirus Disease 2019 (COVID-19) Diagnosis Using the Rational Selection of Serological Biomarkers
title_short	Development of an Enzyme-Linked Immunosorbent Assay (ELISA) for Accurate and Prompt Coronavirus Disease 2019 (COVID-19) Diagnosis Using the Rational Selection of Serological Biomarkers
title_full	Development of an Enzyme-Linked Immunosorbent Assay (ELISA) for Accurate and Prompt Coronavirus Disease 2019 (COVID-19) Diagnosis Using the Rational Selection of Serological Biomarkers
title_fullStr	Development of an Enzyme-Linked Immunosorbent Assay (ELISA) for Accurate and Prompt Coronavirus Disease 2019 (COVID-19) Diagnosis Using the Rational Selection of Serological Biomarkers
title_full_unstemmed	Development of an Enzyme-Linked Immunosorbent Assay (ELISA) for Accurate and Prompt Coronavirus Disease 2019 (COVID-19) Diagnosis Using the Rational Selection of Serological Biomarkers
title_sort	development of an enzyme-linked immunosorbent assay (elisa) for accurate and prompt coronavirus disease 2019 (covid-19) diagnosis using the rational selection of serological biomarkers
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/f029449ab9494f3884ccbea4c8e15902
work_keys_str_mv	AT theanolagousi developmentofanenzymelinkedimmunosorbentassayelisaforaccurateandpromptcoronavirusdisease2019covid19diagnosisusingtherationalselectionofserologicalbiomarkers AT johnroutsias developmentofanenzymelinkedimmunosorbentassayelisaforaccurateandpromptcoronavirusdisease2019covid19diagnosisusingtherationalselectionofserologicalbiomarkers AT vanaspoulou developmentofanenzymelinkedimmunosorbentassayelisaforaccurateandpromptcoronavirusdisease2019covid19diagnosisusingtherationalselectionofserologicalbiomarkers
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Development of an Enzyme-Linked Immunosorbent Assay (ELISA) for Accurate and Prompt Coronavirus Disease 2019 (COVID-19) Diagnosis Using the Rational Selection of Serological Biomarkers

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