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A 10-Minute “Mix and Read” Antibody Assay for SARS-CoV-2

Accurate and rapid diagnostic tools are needed for management of the ongoing coronavirus disease 2019 (COVID-19) pandemic. Antibody tests enable detection of individuals past the initial phase of infection and help examine vaccine responses. The major targets of human antibody response in severe acu...

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Autores principales: Rusanen, Juuso, Kareinen, Lauri, Levanov, Lev, Mero, Sointu, Pakkanen, Sari H., Kantele, Anu, Amanat, Fatima, Krammer, Florian, Hedman, Klaus, Vapalahti, Olli, Hepojoki, Jussi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7908974/
https://www.ncbi.nlm.nih.gov/pubmed/33498157
http://dx.doi.org/10.3390/v13020143
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author Rusanen, Juuso
Kareinen, Lauri
Levanov, Lev
Mero, Sointu
Pakkanen, Sari H.
Kantele, Anu
Amanat, Fatima
Krammer, Florian
Hedman, Klaus
Vapalahti, Olli
Hepojoki, Jussi
author_facet Rusanen, Juuso
Kareinen, Lauri
Levanov, Lev
Mero, Sointu
Pakkanen, Sari H.
Kantele, Anu
Amanat, Fatima
Krammer, Florian
Hedman, Klaus
Vapalahti, Olli
Hepojoki, Jussi
author_sort Rusanen, Juuso
collection PubMed
description Accurate and rapid diagnostic tools are needed for management of the ongoing coronavirus disease 2019 (COVID-19) pandemic. Antibody tests enable detection of individuals past the initial phase of infection and help examine vaccine responses. The major targets of human antibody response in severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are the spike glycoprotein (SP) and nucleocapsid protein (NP). We have developed a rapid homogenous approach for antibody detection termed LFRET (protein L-based time-resolved Förster resonance energy transfer immunoassay). In LFRET, fluorophore-labeled protein L and antigen are brought to close proximity by antigen-specific patient immunoglobulins of any isotype, resulting in TR-FRET signal. We set up LFRET assays for antibodies against SP and NP and evaluated their diagnostic performance using a panel of 77 serum/plasma samples from 44 individuals with COVID-19 and 52 negative controls. Moreover, using a previously described SP and a novel NP construct, we set up enzyme linked immunosorbent assays (ELISAs) for antibodies against SARS-CoV-2 SP and NP. We then compared the LFRET assays with these ELISAs and with a SARS-CoV-2 microneutralization test (MNT). We found the LFRET assays to parallel ELISAs in sensitivity (90–95% vs. 90–100%) and specificity (100% vs. 94–100%). In identifying individuals with or without a detectable neutralizing antibody response, LFRET outperformed ELISA in specificity (91–96% vs. 82–87%), while demonstrating an equal sensitivity (98%). In conclusion, this study demonstrates the applicability of LFRET, a 10-min “mix and read” assay, to detection of SARS-CoV-2 antibodies.
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spelling pubmed-79089742021-02-27 A 10-Minute “Mix and Read” Antibody Assay for SARS-CoV-2 Rusanen, Juuso Kareinen, Lauri Levanov, Lev Mero, Sointu Pakkanen, Sari H. Kantele, Anu Amanat, Fatima Krammer, Florian Hedman, Klaus Vapalahti, Olli Hepojoki, Jussi Viruses Article Accurate and rapid diagnostic tools are needed for management of the ongoing coronavirus disease 2019 (COVID-19) pandemic. Antibody tests enable detection of individuals past the initial phase of infection and help examine vaccine responses. The major targets of human antibody response in severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are the spike glycoprotein (SP) and nucleocapsid protein (NP). We have developed a rapid homogenous approach for antibody detection termed LFRET (protein L-based time-resolved Förster resonance energy transfer immunoassay). In LFRET, fluorophore-labeled protein L and antigen are brought to close proximity by antigen-specific patient immunoglobulins of any isotype, resulting in TR-FRET signal. We set up LFRET assays for antibodies against SP and NP and evaluated their diagnostic performance using a panel of 77 serum/plasma samples from 44 individuals with COVID-19 and 52 negative controls. Moreover, using a previously described SP and a novel NP construct, we set up enzyme linked immunosorbent assays (ELISAs) for antibodies against SARS-CoV-2 SP and NP. We then compared the LFRET assays with these ELISAs and with a SARS-CoV-2 microneutralization test (MNT). We found the LFRET assays to parallel ELISAs in sensitivity (90–95% vs. 90–100%) and specificity (100% vs. 94–100%). In identifying individuals with or without a detectable neutralizing antibody response, LFRET outperformed ELISA in specificity (91–96% vs. 82–87%), while demonstrating an equal sensitivity (98%). In conclusion, this study demonstrates the applicability of LFRET, a 10-min “mix and read” assay, to detection of SARS-CoV-2 antibodies. MDPI 2021-01-20 /pmc/articles/PMC7908974/ /pubmed/33498157 http://dx.doi.org/10.3390/v13020143 Text en © 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Rusanen, Juuso
Kareinen, Lauri
Levanov, Lev
Mero, Sointu
Pakkanen, Sari H.
Kantele, Anu
Amanat, Fatima
Krammer, Florian
Hedman, Klaus
Vapalahti, Olli
Hepojoki, Jussi
A 10-Minute “Mix and Read” Antibody Assay for SARS-CoV-2
title A 10-Minute “Mix and Read” Antibody Assay for SARS-CoV-2
title_full A 10-Minute “Mix and Read” Antibody Assay for SARS-CoV-2
title_fullStr A 10-Minute “Mix and Read” Antibody Assay for SARS-CoV-2
title_full_unstemmed A 10-Minute “Mix and Read” Antibody Assay for SARS-CoV-2
title_short A 10-Minute “Mix and Read” Antibody Assay for SARS-CoV-2
title_sort 10-minute “mix and read” antibody assay for sars-cov-2
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7908974/
https://www.ncbi.nlm.nih.gov/pubmed/33498157
http://dx.doi.org/10.3390/v13020143
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