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.
Keywords: CBA functional beads; COVID-19; IgG; IgM and IgA antibodies; SARS-CoV-2; flow cytometry; multiplex immunoassay.
【저자키워드】 COVID-19, SARS-CoV-2, IgG, flow cytometry, CBA functional beads, IgM and IgA antibodies, multiplex immunoassay., 【초록키워드】 IgM, antibody, Diagnosis, flow cytometry, Protein, immunoassay, sensitivity, specificity, Immunoglobulin G, SARS-CoV-2 antibodies, Immunoglobulin, IgA, multiplex, Cytometry, Flow, opens, IgA antibodies, positive, SARS-CoV-2 nucleocapsid, secondary antibodies, IgA2, anti-IgG, anti-IgM, streptavidin, functional, condition, were used, biotinylated, CBA, COVID-19 negative, 【제목키워드】 IgM, nucleocapsid protein, anti-SARS-CoV-2 IgG, IgA, approach, flow cytometric,