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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 (2021). A 10-Minute “Mix and Read” Antibody Assay for SARS-CoV-2. Viruses, 13(2):143.

Abstract

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.

Abstract

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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Additional indexing

Item Type:Journal Article, refereed, original work
Communities & Collections:05 Vetsuisse Faculty > Veterinärwissenschaftliches Institut > Institute of Veterinary Pathology
Dewey Decimal Classification:570 Life sciences; biology
Scopus Subject Areas:Health Sciences > Infectious Diseases
Life Sciences > Virology
Uncontrolled Keywords:Virology, Infectious Diseases
Language:English
Date:20 January 2021
Deposited On:22 Nov 2021 11:41
Last Modified:26 Jun 2024 01:43
Publisher:MDPI Publishing
ISSN:1999-4915
OA Status:Gold
Free access at:Publisher DOI. An embargo period may apply.
Publisher DOI:https://doi.org/10.3390/v13020143
Project Information:
  • : FunderSNSF
  • : Grant ID1000-000001
  • : Project TitleSchlussband (Bd. VI) der Jacob Burckhardt-Biographie
  • : FunderSNSF
  • : Grant ID1000-000001
  • : Project TitleSchlussband (Bd. VI) der Jacob Burckhardt-Biographie
  • : FunderSNSF
  • : Grant ID1000-000001
  • : Project TitleSchlussband (Bd. VI) der Jacob Burckhardt-Biographie
  • : FunderSNSF
  • : Grant ID1000-000001
  • : Project TitleSchlussband (Bd. VI) der Jacob Burckhardt-Biographie
  • : FunderSNSF
  • : Grant ID1000-000001
  • : Project TitleSchlussband (Bd. VI) der Jacob Burckhardt-Biographie
  • : FunderFP7
  • : Grant ID234147
  • : Project TitleGLFEM - Generic Linking of Finite Element based Models
  • : FunderSNSF
  • : Grant ID1000-000001
  • : Project TitleSchlussband (Bd. VI) der Jacob Burckhardt-Biographie
  • : FunderSNSF
  • : Grant ID1000-000001
  • : Project TitleSchlussband (Bd. VI) der Jacob Burckhardt-Biographie
  • Content: Published Version
  • Language: English
  • Licence: Creative Commons: Attribution 4.0 International (CC BY 4.0)