SARS-CoV-2 Human IgG Quantitative (4-Plex) Multiplex ELISA Kit (For RUO purposes)

ELISA kit for Multiplex detection

Attention: this product requires a special plate reader. Please make sure you have the proper equipment before ordering. For details please contact [email protected]

Product Info Summary

Species Human
Assay Type Indirect
Sample Type Human Serum, Plasma
Assay Length 2 hours
Specificity See Assay Validation Below
Volume Required Per Well Min 2µl
Detection Method Chemiluminescent
Multiplex Format 96-well solid plate
SKU MEK2009-2
Manufacturer Reference Number 711649HU

Product Overview

The Q-Plex SARS-CoV-2 Human IgG Quantitative (4-plex) Multiplex ELISA Kit is a quantitative chemiluminescent assay (ELISA) allowing concurrent measurement of human IgG antibodies to SARS-CoV-2 S1 and SARS-CoV-2 S2 proteins in serum and plasma samples. Sensitive, reproducible, and convenient 96-well format requiring low sample volumes. Requires the Q-View™ Imaging System for analysis.

Assay Principle

The Q-Plex SARS-CoV-2 Human IgG Quantitative (4-Plex) multiplex assay is a quantitative enzyme-linked immunosorbent assay that detects IgG antibodies to SARS-CoV-2 in human serum or plasma. SARS-CoV-2 is the novel coronavirus that causes COVID-19. The SARS-CoV-2 virus has several structural proteins including two spike proteins, S1 and S2. When an individual is infected with the SARS-CoV-2 virus, their immune system produces antibodies to these viral proteins. The typical immune response produces detectable antibody levels ~8-10 days following the onset of symptoms.

The Q-Plex SARS-CoV-2 Human IgG Quantitative (4-Plex) Multiplex ELISA measures IgG antibodies to both the S1 and S2 spike proteins present in a blood sample. The assay only quantifies IgG antibodies, not other classes of antibodies such as IgM or IgA.

Samples or controls are pipetted into wells of an arrayed microplate thereby immobilizing antibodies to SARS-CoV-2 S1, SARS-CoV-2 S2, Sheep Fc (negative control), and Human IgGs (positive control) to their locations in the array. After washing away any unbound IgG, a mixture that contains biotinylated anti-human IgG is added. After washing away unbound biotinylated antibody, streptavidin-horseradish peroxidase (SHRP) is added. Following an additional wash, the amount of SHRP remaining on each location of the array is proportional to the amount of Human IgG antibody reactive to SARS-Cov2 S1, SARS-Cov2 S2, Sheep Fc (negative control), anti-Human IgG (positive control) initially captured.

The amount of conjugated enzyme on each location of the array is measured with the addition of a chemiluminescent substrate.

The multiplex assay allows for simultaneous indirect ELISA on the following four printed spots:

  1. SARS-CoV-2 Spike Glycoprotein (S1), a recombinant antigen which contains amino acids 1-674 of subunit 1. Spike S1 is expressed in mammalian HEK293 cells with a Sheep Fc-Tag.
  2. SARS-CoV-2 Spike Glycoprotein (S2) is a recombinant antigen which contains the Spike subunit 2 protein, amino acids 685-1211. Spike S2 is expressed in mammalian HEK293 cells with a Sheep Fc-Tag.
  3. Sheep Fc is a negative control to ensure no cross-reactivity occurs between human IgGs in the sample and the Fc-Tag on the SARS-Cov-2 Spike proteins.
  4. Anti-Human IgG is a positive control to ensure the kit performs and the IFU was followed correctly.

Assay Validation

The Q-Plex SARS-CoV-2 Human IgG Quantitative (4-Plex) ELISA tests for IgG antibodies to either the SARS-CoV-2 S1 protein or the SARS-CoV-2 S2 protein. Our validation studies identified measurable improvement in clinical performance when results from the S1 and S2 assays are considered together rather than single assays (see assay validation data below). Quansys recommends that a sample should only be considered positive for the presence of SARS-CoV-2 reactive IgGs when antibodies reactive to both S1 and S2 are detected.

Negative samples, collected prior to August 2019, and known positive samples collected from individuals who tested positive for COVID-19 on a molecular test at least 14 days prior to sample collection were used to determine clinical sensitivity and specificity.

Evaluating the S1 and S2 simultaneously allows for greater clinical sensitivity and specificity.

SARS-CoV-2 IgG Assay vs. Molecular COVID-19 Test
N = 254 Confirmed Positive Confirmed Negative
IgG Test Positive 33 0
IgG Test Negative 1 220
The combined S1 & S2 reactive antibody assay demonstrates:
Estimated Sensitivity (PPA) 97%
Estimated Specificity (NPA) 100%
PPV 100%
NPV 100%
SARS-CoV-2 IgG Assay Characteristics
Principle Indirect ELISA
Sample Type Serum, Plasma, Whole Blood
Sample Volume 2 µL
Assay Incubation 2 hours, RT
Total Wash Steps 3
Within Plate Reproducibility 8% CV
Between Plate Reproducibility 10% CV

Kit Components

Each kit contains a 96-well plate, featuring the relevant biomarker panel in each well, and all reagents required to perform testing.

Reagents Include:

  • Detection Mix
  • Controls
  • Substrate
  • Sample Diluents
  • Streptavidin HRP
  • Wash Buffer

Kit Processing

The Multiplex assays require the use of the Q-View™ software to read and interpret the test results. A fully-functional, free trial version of the software is available to download, install, and use to analyze your first kit (s). At the end of the trial period, a purchased license is required to continue the use of the Q-View software.

Q-Plex arrays are developed and optimized to work with Q-View imagers. We do not guarantee the results obtained from other imagers because not all imagers are compatible with Q-Plex arrays.

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MEK2009-2
The multiplex ELISA kits require special plate readers. For more info see special plate reader. The $800 demo option is for new customer only, see more info at demo options.
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