Multiplexed flow cytometric approach for detection of anti-SARS-CoV-2 IgG,IgM and IgA using beads covalently coupled to the nucleocapsid protein |
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| Authors: | IF Zattoni LF Huergo ECM Gerhardt JM Nardin AMF dos Santos FGM Rego G Picheth VR Moure G Valdameri |
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| Institution: | 1. Pharmaceutical Sciences Graduate Program, Laboratory of Cancer Drug Resistance, Federal University of Paraná, Curitiba, PR, Brazil;2. Setor Litoral, Federal University of Paraná, Matinhos, PR, Brazil;3. Department of Biochemistry and Molecular Biology, Federal University of Paraná, Curitiba, PR, Brazil;4. Hospital Erasto Gaertner, Curitiba, PR, Brazil;5. Department of Clinical Analysis, Federal University of Paraná, Curitiba, PR, Brazil |
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| Abstract: | 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. |
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| Keywords: | CBA functional beads COVID-19 flow cytometry IgG IgM and IgA antibodies multiplex immunoassay SARS-CoV-2 |
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