The largest database of trusted experimental protocols

Quan t cell sars cov 2

Manufactured by EUROIMMUN
Sourced in Germany

The Quan-T-Cell SARS-CoV-2 is a laboratory equipment product designed to quantify SARS-CoV-2-specific T-cell responses. It provides an in vitro functional assay to measure the activation of SARS-CoV-2-specific T cells upon stimulation with viral antigens.

Automatically generated - may contain errors

Lab products found in correlation

7 protocols using quan t cell sars cov 2

1

SARS-CoV-2 Spike-Specific T Cell Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols
The IFN-γ and chemokine release assay “Quan-T-Cell SARS-CoV-2” (Euroimmun, Germany) was used to detect spike S1-specific T cells. For logistic reasons, we performed the assay for infection-naïve vaccinated individuals with PBMC and for SARS-CoV-2-infected individuals with heparin whole blood samples. The assay was conducted with three tubes, an unstimulated control (BLANK), an S1-coated tube (TUBE), and a mitogen-coated positive control (STIM). To compare PBMC and whole blood stimulations, we calculated the required cell number for PBMC stimulation based on Trucount™ analyses of 500 µl whole blood samples (mean values). Moreover, we did not compare absolute concentrations of secreted cytokines, instead we used the fold induction which was calculated for each patient individually (TUBE/BLANK). PBMC were thawed and seeded with 7×105 to 1×106 per tube in 500 µl medium (RPMI1640 +2mM L-Glutamine + 100 U/mL Penicillin-Streptomycin +1mM sodium pyruvate +10% FCS) or 500 µl heparin-whole blood was added to each tube. Subsequently, tubes were inverted and incubated for 20 h at 37°C with 5% CO2. After the incubation, supernatant or plasma was collected by centrifuging the tube for 10 min at 6000g. The supernatant and plasma were frozen until further usage. Cytokines and chemokines in the supernatant or plasma were measured via Luminex-based multiplex assays.
+ Open protocol
+ Expand
2

SARS-CoV-2 Spike Protein T Cell Response

Check if the same lab product or an alternative is used in the 5 most similar protocols
The spike protein-specific T cell response was determined using a SARS-CoV-2 interferon (IFN)-γ release assay (IGRA) kit (Quan-T-Cell SARS-CoV-2, Euroimmun Medizinische Labordiagnostika, Luebeck, Germany). Whole blood collected in lithium heparin tubes was stimulated in blank, spike protein-, and mitogen-coated tubes for 16 h. The samples were then centrifuged to isolate and freeze the stimulated plasma, which was then subjected to an IFN-γ assay by ELISA according to the manufacturer’s protocol. The T cell response capacity was determined by subtracting the IFN-γ concentration (mIU/mL) in the test tube from that in the blank tube. According to the manufacturer, a value of >100 was regarded as a detectable response.
+ Open protocol
+ Expand
3

Quantifying T-Cell Response to SARS-CoV-2

Check if the same lab product or an alternative is used in the 5 most similar protocols
To evaluate cellular response against viral proteins Quan-T-Cell SARS-CoV-2 (Euroimmun, Lübeck, Germany) test was used. A cut-off value of 18.44 mIU/mL was used to define positive test values according to the manufacturer’s instructions. In the first stage, freshly drawn heparinized whole blood was incubated with the S1 antigen of the SARS-CoV-2 virus coated on the bottom of the test tube. Whole blood was also incubated in a second negative control tube (assessment of non-specific background response) and a third positive control tube (assessment of overall T cell response after stimulation). After incubation time (22-24h), serum plasma was obtained. In the second stage, an ELISA test was performed to measure the secreted IFN-γ in the first step of the test.
+ Open protocol
+ Expand
4

SARS-CoV-2 Spike Protein T-Cell Response

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cellular immune response was assessed with IGRA. A specific stimulation of T-cells by the spike protein of SARS-CoV-2 was performed using the Quan-T-Cell SARS-CoV-2 (Euroimmun AG, Lübeck, Germany) to determine the amount of IFN-γ released by immune cells. IFN-y responses were then measured using the Quan-T-Cell ELISA (Euroimmun AG, Lübeck, Germany) Interferon Gamma Release Assay. Results were obtained in milli-international units per milliliter (mIU/mL) in accordance with the manufacturer’s instructions. Values below 100 mIU/mL were interpreted as “IGRA-negative,” and values above 200 mIU/mL were interpreted as “IGRA-positive.”
+ Open protocol
+ Expand
5

Cellular Immunity Measurement for SARS-CoV-2

Check if the same lab product or an alternative is used in the 5 most similar protocols
As described in our previous study, for the analysis of cellular immunity, the Quan-T-Cell SARS-CoV-2 in combination with the Quan-T-Cell ELISA (Euroimmun Medizinische Labordiagnostika, Lübeck, Germany) was used [15 (link)]. The principle of the test is a measurement of IFN-γ concentration released by activated immune cells. Fresh whole blood samples were collected in heparinized tubes and pipetted into the three stimulation tubes (Quan-T-Cell SARS-CoV-2): (1) COV-2 IGRA (interferon-gamma release assay) Blank was used for measuring individual IFN-γ concentrations as it contained no activating components; (2) CoV-2 IGRA Tube was coated with peptide components of the S1 domain of the SARS-CoV-2 spike protein; and (3) CoV-2 IGRA Stim was coated with mitogen to verify if the sample contained a sufficient number of viable and functional T cells. After incubation of the individual whole blood in the stimulation tubes for 20–24 h at 37 °C, the separated plasma was used to determine IFN-γ concentration by Quan-T-Cell ELISA.
+ Open protocol
+ Expand
6

Determining Cellular Immune Responses after Vaccination

Check if the same lab product or an alternative is used in the 5 most similar protocols
For the determination of cellular immune responses after two-dose vaccination, an interferon gamma (IFNγ) release assay (IGRA) (Quan-T-Cell SARS-CoV-2 & Quan-T-Cell-ELISA, Euroimmun, Lübeck, Germany) was applied as previously described [7] (link). A cut-off of 100 mIE/ml was defined as a positive IFNγ release response.
+ Open protocol
+ Expand
7

Quantitative SARS-CoV-2 T-Cell IFN-γ Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols
The Quan-T-Cell System combine the Quan-T-Cell ELISA and the Quan-T-Cell SARS-CoV-2 (Euroimmun, Lübeck, Germany) for the quantitative determination of IFN-γ released by T cells specific for SARS-CoV-2. Briefly, 0.5 mL of fresh heparinized whole blood was added to 3 test tubes: (1) CoV-2 IGRA BLANK: no T-cell stimulation, for assess of the IFN-γ background; (2) CoV-2 IGRA TUBE: specific T-cell stimulation by antigens of SARS-CoV-2 spike protein; (3) CoV-2 IGRA STIM: unspecific T-cell stimulate on by means of a mitogen, for control of the stimulation ability. After sampling, the test tubes were inverted six times and incubated for 16 to 20 hours at 37°C. The samples were then centrifuged at 12000 RCF for 10 minutes and the plasma supernatant was transferred into an eppendorf and stored at– 20°C until testing. IFN-γ was finally detected in the supernatants by an enzyme-linked immunosorbent assay (ELISA) using the Euroimmun Analyzer I instrument (Euroimmun, Lübeck, Germany). IFN-γ response was measured by subtracting the result in the stimulated tube minus the result in the unstimulated tube. The test validation criteria were as follows: IFN-γ [blank] < 400 mIU/mL and IFN-γ [stim]–IFN-γ [blank] > 400 mIU/mL. Results were interpreted as follows: IFN-γ [SARS-CoV-2]–IFN-γ [blank] < 100 mIU/mL negative, 100–200 mIU/mL borderline, and > 200 mIU/mL as positive.
+ Open protocol
+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!