This test requires professional lab infrastructure and staff and must be ordered and administered by medical professionals. The test cannot be performed by private individuals and we do not sell the kits to such subjects. If you suspect you are infected with SARS-CoV-2, please contact your healthcare provider.

The Cov19 FluoBoltTM-DAT allows the simultaneous and quantitative measurement of antibodies against the S1RBD AND nucleocapsid antigen of SARS-CoV-2 within one single measurement by using only 10 µl of sample and generates results within 60 minutes.

In contrast to other antibody assays, which detect all antibodies generated against the S1- or NC-protein, the Cov19 FluoBoltTM-DAT is epitope specific and detects immune-dominant antibodies species, i.e. the most important ones generated by either vaccination or infection. This is also nicely demonstrated by the fact, that it detects anti-S1RBD against all dominant virus variants identified so far (see chart in specification tab).

The calibrators of this assay allow for a concentration (µg/mL) calculation of antibodies in a sample.  Therefore, the Cov19 FluoBoltTM-DAT assay may be valuable tool to establish an antibody based “protective correlate”, i.e. what amount and type of antibody gives protection for a certain period of time.  The assay is also a valuable tool to differentiate unvaccinated vs only vaccinated vs infected samples in any SARS-CoV-2 research study.

 

How the Cov19 FluoBoltTM-DAT test works:

When performing the assay, 10 µl of a serum or plasma sample are simultaneously mixed with 50 µl of two different fluorescence-labeled tracer antibodies against the S1RBD and against the NC protein and incubated in FluoBoltTM‘s special fluorescence-enhancing microtiter plate for 60 minutes. By measuring at 2 different wavelengths, their displacement by homologous antibodies from the sample is determined simultaneously. The signals at one wavelength provide information about the concentration of anti-S1RBD antibodies. The signals at the other wavelength indicate the concentration of anti-nucleocapsid antibodies.

By using the supplied calibrators, a calibration curve for the anti-S1RBD and the anti-NC antibodies is constructed, and the antibody concentration of a sample is read from it.

With just a single measurement, this results in a quantitative determination of these two antibody species against SARS-CoV-2, which represent most of the immune response and may protect against infection and illness.

For measurement, any commercially available fluorescence microtiter plate reader can be used.

• 1 x 96 well- MEF-microtiter plate pre-coated with SARS-CoV-2 NC/S1RBD fusion antigen
• 1 x 25 ml- Wash buffer concentrate 20x, natural cap
• 1 x 5 ml- Antibody tracer mixture consisting of FITC labelled anti-NC antibody and Cy5 labelled anti-S1RBD antibody in black vial with black cap
• 5 vials x 0.1 ml- Antibody standard mixture consisting of anti-NC antibodies and anti-S1RBD antibodies in human serum (40, 20, 10, 5, and 0 ug/mL), provided lyophilized in glass vials with white caps
• 2 vials- Antibody controls (A with yellow cap and B with green cap). A is the HIGH control and B is the LOW control.
• 1 x 10 ml- Ready to use sample diluent in plastic vial with natural cap
• 1 x 96 well plate for tracer and sample pre-mixing

ADDITIONAL MATERIAL SUPPLIED WITH THE KIT:
2 self-adhesive plastic films
QC data sheet
Protocol sheet
Instruction manual for use
2 desiccant bags for plate storage

MATERIAL AND EQUIPMENT REQUIRED BUT NOT SUPPLIED:
Precision pipettes calibrated to deliver 10 μl, 20 μl, 50 μl, 200 μl, 500 μl, and disposable tips
Distilled or deionized water
Plate washer, multichannel pipette, or manifold dispenser for washing
Refrigerator to keep kit at 4°C (2-8°C)
Fluorescence microplate reader

Method Metal Enhanced Direct Sandwich Fluorescence Immunoassay in 96-well format
Sample Type Serum and Plasma
Standard Range 0 to 40 µg/mL

5 standards and 2 controls in a serum based matrix

Sample Volume 10 µL (undiluted sample)/well
Incubation/time/temperature Single-step assay / 60 min / Room Temperature
Sensitivity LOD 0.1 µg/mL for both antibodies
Specificity Negative Agreement 100%

Positive Agreement 95%

Anti-S1RBD results correlated very well with the results from the surrogate virus neutralization test, which is good support that the assay indeed detects neutralizing antibodies.

Measuring the response to the fluorescent labeled NC-antibody tracer did only show binding to the fusion wild-type spike protein as expected. This is of course not very surprising, since the S1RBD variants did not have any part of the NC-protein. However, the kit detects anti-S1RBD against all dominant virus variants identified so far.

Marketing Literature:  FluoBolt Cov19-DAT Flyer