Details

Autor(en) / Beteiligte

• Zattoni, I.F.
• Huergo, L.F.
• Gerhardt, E.C.M.
• Nardin, J.M.
• Santos, A.M.F.
• Rego, F.G.M.
• Picheth, G.
• Moure, V.R.
• Valdameri, G.

Titel

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

Ist Teil von

• Letters in applied microbiology, 2022-06, Vol.74 (6), p.863-872

Ort / Verlag

England: Oxford University Press

Erscheinungsjahr

2022

Link zum Volltext

Quelle

Wiley Blackwell Single Titles

Beschreibungen/Notizen

• 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. Significance and Impact of the Study: Flow cytometry has emerged as a promising technique for detection of anti‐SARS‐CoV‐2 antibodies. In this study, we developed an innovative strategy for simultaneous detection of immunoglobulin G (IgG), IgM and IgA. This new cell‐free assay efficiently discriminate COVID‐19 negative and positive samples. The simultaneous detection of IgG, IgM and IgA showed a high sensitivity and specificity. This novel strategy opens a new avenue for flow cytometry‐based diagnosis.