This ELISA kit uses the Sandwich-Ag ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with recombinant SARS-CoV-2 Spike Protein. After adding samples (or controls) to wells, the total antibodies against SARS-CoV-2 Spike Protein will combine with the pre-coated SARS-CoV-2 Spike Protein. After washing completely, add Horseradish Peroxidase (HRP) conjugated SARS-CoV-2 Spike Protein to develop the antigen-antibody-HRP conjugated antigen complex. Free components are washed away. The substrate solution is added to each well. Only those wells that contain SARS-CoV-2 Spike Protein, antibodies against SARS-CoV-2 Spike Protein and HRP conjugated SARS-CoV-2 Spike Protein will appear blue in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 ± 2 nm. Compared with the CUT OFF value to judge whether SARS-CoV-2 Spike Protein total antibody exists in the tested samples or not.

SARS-CoV-2, which causes the global pandemic coronavirus disease 2019 (Covid-19), belongs to a family of viruses known as coronaviruses that are commonly comprised of four structural proteins: Spike protein (S), Envelope protein (E), Membrane protein (M), and Nucleocapsid protein (N) . SARS-CoV-2 Spike Protein (S Protein) is a glycoprotein that mediates membrane fusion and viral entry. The S protein is homotrimeric, with each ~180-kDa monomer consisting of two subunits, S1 and S2. In SARS-CoV-2, as with most coronaviruses, proteolytic cleavage of the S protein into the S1 and S2 subunits is required for activation. The S1 subunit is focused on attachment of the protein to the host receptor while the S2 subunit is involved with cell fusion. The S protein of SARS-CoV-2 shares 75% and 29% amino acid (aa) sequence identity with the S protein of SARS-CoV-1 and MERS, respectively.The S Protein of the SARS-CoV-2 virus, like the SARS-CoV-1 counterpart, binds Angiotensin-Converting Enzyme 2 (ACE2), but with much higher affinity and faster binding kinetics through the receptor binding domain (RBD) located in the C-terminal region of S1. Based on structural biology studies, the RBD can be oriented either in the up无standing or down无lying state with the up无standing state associated with higher pathogenicity. Polyclonal antibodies to the RBD of the SARS-CoV-2 protein have been shown to inhibit interaction with the ACE2 receptor, confirming RBD as an attractive target for vaccinations or antiviral therapy. It has been demonstrated that the S Protein can invade host cells through the CD147无EMMPRIN receptor and mediate membrane fusion. A SARS-CoV-2 variant carrying the S protein aa change D614G has become the most prevalent form in the global pandemic and has been associated with greater infectivity and higher viral load .