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Journal of Clinical Microbiology
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Immunoassays

Development and validation of a multiplex microsphere immunoassay using dried blood spots for SARS-CoV-2 seroprevalence: application in first responders in Colorado, USA

Jonathan S. Schultz, Mary K. McCarthy, Cody Rester, Katherine R. Sabourin, Kyle Annen, Melkon DomBourian, Elan Eisenmesser, Ashely Frazer-Abel, Vijaya Knight, Thomas Jaenisch, Thomas E. Morrison, Rosemary Rochford, Ross M. Kedl
Jonathan S. Schultz
a Division of Infectious Diseases, School of Medicine, University of Colorado, Anschutz Medical Campus, Aurora, CO, USA
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  • ORCID record for Jonathan S. Schultz
Mary K. McCarthy
b Department of Immunology and Microbiology, University of Colorado, Anschutz Medical Campus, Aurora, CO, USA
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Cody Rester
b Department of Immunology and Microbiology, University of Colorado, Anschutz Medical Campus, Aurora, CO, USA
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Katherine R. Sabourin
b Department of Immunology and Microbiology, University of Colorado, Anschutz Medical Campus, Aurora, CO, USA
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Kyle Annen
c Clinical & Translational Allergy and Immunology Laboratory, Children’s Hospital of Colorado, University of Colorado, Anschutz Medical Campus, Aurora, CO, USA
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Melkon DomBourian
c Clinical & Translational Allergy and Immunology Laboratory, Children’s Hospital of Colorado, University of Colorado, Anschutz Medical Campus, Aurora, CO, USA
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Elan Eisenmesser
d Department of Biochemistry and Molecular Genetics, School of Medicine, University of Colorado, Anschutz Medical Campus, Aurora, CO, USA
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Ashely Frazer-Abel
e Exsera Biolabs, Department of Rheumatology, University of Colorado, Anschutz Medical Campus, Aurora, CO, USA
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Vijaya Knight
c Clinical & Translational Allergy and Immunology Laboratory, Children’s Hospital of Colorado, University of Colorado, Anschutz Medical Campus, Aurora, CO, USA
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Thomas Jaenisch
f Department of Epidemiology, Colorado School of Public Health, Anschutz Medical Campus, Aurora, CO, USA
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Thomas E. Morrison
b Department of Immunology and Microbiology, University of Colorado, Anschutz Medical Campus, Aurora, CO, USA
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Rosemary Rochford
b Department of Immunology and Microbiology, University of Colorado, Anschutz Medical Campus, Aurora, CO, USA
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  • For correspondence: Rosemary.Rochford@cuanschutz.edu
Ross M. Kedl
b Department of Immunology and Microbiology, University of Colorado, Anschutz Medical Campus, Aurora, CO, USA
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DOI: 10.1128/JCM.00290-21
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ABSTRACT

Serological testing of large representative populations for antibodies to SARS-CoV-2 is needed to estimate seroprevalence, transmission dynamics, and the duration of antibody responses from natural infection and vaccination. In this study, a high-throughput SARS-CoV-2 multiplex microsphere immunoassay (MMIA) was developed for the receptor binding domain (RBD) and nucleocapsid (N) that was more sensitive than ELISA (98% vs 87%). The MMIA was then applied and validated in 264 first responders in Colorado using serum and dried blood spot (DBS) eluates, compared to ELISA and evaluated for neutralizing antibodies. Four percent (11/264) of first responders were seropositive in July-August 2020. Serum and DBS were highly correlated for anti-RBD and anti-N antibodies (R=0.83, p<0.0001 and R=0.87, p<0.0001, respectively) by MMIA. The MMIA accurately predicted SARS-CoV-2 neutralizing antibodies using DBS (R=0.76, p=0.037). On repeat antibody testing three months later, anti-RBD IgG decreased less rapidly than anti-N IgG measured by MMIA, with a median change in gMFI of 62% vs 79% (p<0.01), for anti-RBD and anti-N IgG respectively. This novel MMIA using DBS could be scalable for rapid and affordable SARS-CoV-2 serosurveillance in the U.S. and globally.

  • Copyright © 2021 American Society for Microbiology.

All Rights Reserved.

This article is made available via the PMC Open Access Subset for unrestricted noncommercial re-use and secondary analysis in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic.

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Development and validation of a multiplex microsphere immunoassay using dried blood spots for SARS-CoV-2 seroprevalence: application in first responders in Colorado, USA
Jonathan S. Schultz, Mary K. McCarthy, Cody Rester, Katherine R. Sabourin, Kyle Annen, Melkon DomBourian, Elan Eisenmesser, Ashely Frazer-Abel, Vijaya Knight, Thomas Jaenisch, Thomas E. Morrison, Rosemary Rochford, Ross M. Kedl
Journal of Clinical Microbiology Apr 2021, JCM.00290-21; DOI: 10.1128/JCM.00290-21

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Development and validation of a multiplex microsphere immunoassay using dried blood spots for SARS-CoV-2 seroprevalence: application in first responders in Colorado, USA
Jonathan S. Schultz, Mary K. McCarthy, Cody Rester, Katherine R. Sabourin, Kyle Annen, Melkon DomBourian, Elan Eisenmesser, Ashely Frazer-Abel, Vijaya Knight, Thomas Jaenisch, Thomas E. Morrison, Rosemary Rochford, Ross M. Kedl
Journal of Clinical Microbiology Apr 2021, JCM.00290-21; DOI: 10.1128/JCM.00290-21
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