SARS-CoV-2 Total Antibody Detection ELISA Kit (Targeting RBD of Spike Proteins)

Principle of the Assay

This assay employs a quantitative sandwich enzyme immunoassay technique and is uniquely suitable for rapid high-throughput detecting the levels of the SARS-CoV-2 total antibody in serum. The recombinant SARS-CoV-2 RBD fragment has been pre-coated onto a microplate. Controls or samples are pipetted into the wells, and then a horseradish peroxidase-conjugated RBD fragment (RBD-HRP) is added to the wells, producing an antigen-antibody-antigen "sandwich complex". Following incubation and wash steps, a substrate is added. A colored product is formed in proportion to the amount of SARS-CoV-2 total antibody present in the sample. The reaction is terminated by the addition of acid and absorbance is measured at 450nm. A positive control is prepared from SARS-CoV-2 RBD antibodies to assure the validity of the results.

Target Information

Coronaviruses are enveloped viruses with a positive-sense RNA genome and with a nucleocapsid of helical symmetry. SARS-CoV-2 has several structural proteins including spike (S), envelope (E), membrane (M), and nucleocapsid (N). The spike protein (S) contains a receptor-binding domain (RBD), which is responsible for recognizing the cell surface receptor, angiotensin-converting enzyme-2 (ACE2). It is found that the RBD of the SARS-CoV-2 S protein strongly interacts with the human ACE2 receptor leading to endocytosis into the host cells of the deep lung and viral replication. This kit is configured and optimized to support research for SARS-CoV-2, it can be used to detect the RBD antibodies in the serum of patients with SARS-CoV-2 infection as early as possible, and serving as a tool to evaluate the immune level of other animals to the SARS-COV-2 RBD.

Materials Supplied

Kit Components 96 Wells Quantity/Size
Aluminum pouches with a Microwell Plate coated with SARS-CoV-2 RBD fragment (8×12)
1 plate
Positive Control 2 vials
Negative Control 2 vials
Recombinant SARS-CoV-2 RBD fragment conjugated to horseradish peroxidase (HRP)
2 vials
Diluent Buffer
1 bottle
Wash Buffer Concentrate 20x (PBS with 1% Tween-20) 1 bottle
Substrate Solution 1 bottle
Stop Solution 1 bottle
Adhesive Films 4 pieces
Product insert 1 copy

Storage

Storage Store at 2 - 8°C

Performance Characteristics

Repeatability The coefficient of variation of both intra-assay and inter-assay was less than 10%.
Specificity This assay is specific to SARS-CoV-2 neutralizing antibodies.
Method Verification To evaluate the sensitivity of the SARS-CoV-2 Total Antibody Detection ELISA Kit, positive serum and negative serum were tested by diluted in serial two-fold steps in Diluent Buffer. The figure(below) shows the results with the SARS-CoV-2 Total Antibody Detection ELISA Kit for tested samples: