Multiplexed flow cytometric approach for detection of anti-SARS-CoV-2 IgG, IgM and IgA using beads covalently coupled to the nucleocapsid protein.


Journal

Letters in applied microbiology
ISSN: 1472-765X
Titre abrégé: Lett Appl Microbiol
Pays: England
ID NLM: 8510094

Informations de publication

Date de publication:
Jun 2022
Historique:
revised: 11 01 2022
received: 28 06 2021
accepted: 13 01 2022
pubmed: 12 2 2022
medline: 21 5 2022
entrez: 11 2 2022
Statut: ppublish

Résumé

Flow cytometry has emerged as a promising technique for detection of SARS-CoV-2 antibodies. In this study, we developed an innovative strategy for simultaneous detection of immunoglobulin G (IgG), IgM and IgA. The SARS-CoV-2 nucleocapsid protein was covalently bound to functional beads surface applying sulpho-SMCC chemistry. BUV395 anti-IgG, BB515 anti-IgM, biotinylated anti-IgA1/IgA2 and BV421 streptavidin were used as fluorophore conjugated secondary antibodies. Serum and antibodies reaction conditions were optimized for each antibody isotype detection and a multiplexed detection assay was developed. This new cell-free assay efficiently discriminate COVID-19 negative and positive samples. The simultaneous detection of IgG, IgM and IgA showed a sensitivity of 88·5-96·2% and specificity of 100%. This novel strategy opens a new avenue for flow cytometry-based diagnosis.

Identifiants

pubmed: 35148433
doi: 10.1111/lam.13674
pmc: PMC9115257
doi:

Substances chimiques

Antibodies, Viral 0
Immunoglobulin A 0
Immunoglobulin G 0
Immunoglobulin M 0
Nucleocapsid Proteins 0

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

863-872

Subventions

Organisme : Conselho Nacional de Desenvolvimento Científico e Tecnológico
ID : 400953/2016-1

Informations de copyright

© 2022 The Society for Applied Microbiology.

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Auteurs

I F Zattoni (IF)

Pharmaceutical Sciences Graduate Program, Laboratory of Cancer Drug Resistance, Federal University of Paraná, Curitiba, PR, Brazil.

L F Huergo (LF)

Setor Litoral, Federal University of Paraná, Matinhos, PR, Brazil.

E C M Gerhardt (ECM)

Department of Biochemistry and Molecular Biology, Federal University of Paraná, Curitiba, PR, Brazil.

J M Nardin (JM)

Hospital Erasto Gaertner, Curitiba, PR, Brazil.

A M F Dos Santos (AMF)

Hospital Erasto Gaertner, Curitiba, PR, Brazil.

F G M Rego (FGM)

Department of Clinical Analysis, Federal University of Paraná, Curitiba, PR, Brazil.

G Picheth (G)

Department of Clinical Analysis, Federal University of Paraná, Curitiba, PR, Brazil.

V R Moure (VR)

Pharmaceutical Sciences Graduate Program, Laboratory of Cancer Drug Resistance, Federal University of Paraná, Curitiba, PR, Brazil.
Department of Clinical Analysis, Federal University of Paraná, Curitiba, PR, Brazil.

G Valdameri (G)

Pharmaceutical Sciences Graduate Program, Laboratory of Cancer Drug Resistance, Federal University of Paraná, Curitiba, PR, Brazil.
Department of Clinical Analysis, Federal University of Paraná, Curitiba, PR, Brazil.

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Classifications MeSH