Cargando…
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...
Autores principales: | , , , , , , , , , , |
---|---|
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 |
_version_ | 1783655829334917120 |
---|---|
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. |
format | Online Article Text |
id | pubmed-7908974 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT rusanenjuuso a10minutemixandreadantibodyassayforsarscov2 AT kareinenlauri a10minutemixandreadantibodyassayforsarscov2 AT levanovlev a10minutemixandreadantibodyassayforsarscov2 AT merosointu a10minutemixandreadantibodyassayforsarscov2 AT pakkanensarih a10minutemixandreadantibodyassayforsarscov2 AT kanteleanu a10minutemixandreadantibodyassayforsarscov2 AT amanatfatima a10minutemixandreadantibodyassayforsarscov2 AT krammerflorian a10minutemixandreadantibodyassayforsarscov2 AT hedmanklaus a10minutemixandreadantibodyassayforsarscov2 AT vapalahtiolli a10minutemixandreadantibodyassayforsarscov2 AT hepojokijussi a10minutemixandreadantibodyassayforsarscov2 AT rusanenjuuso 10minutemixandreadantibodyassayforsarscov2 AT kareinenlauri 10minutemixandreadantibodyassayforsarscov2 AT levanovlev 10minutemixandreadantibodyassayforsarscov2 AT merosointu 10minutemixandreadantibodyassayforsarscov2 AT pakkanensarih 10minutemixandreadantibodyassayforsarscov2 AT kanteleanu 10minutemixandreadantibodyassayforsarscov2 AT amanatfatima 10minutemixandreadantibodyassayforsarscov2 AT krammerflorian 10minutemixandreadantibodyassayforsarscov2 AT hedmanklaus 10minutemixandreadantibodyassayforsarscov2 AT vapalahtiolli 10minutemixandreadantibodyassayforsarscov2 AT hepojokijussi 10minutemixandreadantibodyassayforsarscov2 |