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Mendeley readers
Attention Score in Context
| Title |
Validation of an IFNγ/IL2 FluoroSpot assay for clinical trial monitoring
|
|---|---|
| Published in |
Journal of Translational Medicine, June 2016
|
| DOI | 10.1186/s12967-016-0932-7 |
| Pubmed ID | |
| Authors |
Nina Körber, Uta Behrends, Alexander Hapfelmeier, Ulrike Protzer, Tanja Bauer |
| Abstract |
The FluoroSpot assay, an advancement of the ELISpot assay, enables simultaneous measurement of different analytes secreted at a single-cell level. This allows parallel detection of several cytokines secreted by immune cells upon antigen recognition. Easier standardization, higher sensitivity and reduced labour intensity render FluoroSpot assays an interesting alternative to flow-cytometry based assays for analysis of clinical samples. While the use of immunoassays to study immunological primary and secondary endpoints becomes increasingly attractive, assays used require pre-trial validation. Here we describe the assay validation (precision, specificity and linearity) of a FluoroSpot immunological endpoint assay detecting Interferon γ (IFNγ) and Interleukin 2 (IL2) for use in clinical trial immune monitoring. We validated an IFNγ/IL2 FluoroSpot assay to determine Epstein-Barr virus (EBV)-specific cellular immune responses (IFNγ, IL2 and double positive IFNγ + IL2 responses), using overlapping peptide pools corresponding to EBV-proteins BZLF1 and EBNA3A. Assay validation was performed using cryopreserved PBMC of 16 EBV-seropositive and 6 EBV-seronegative donors. Precision was assessed by (i) testing 16 donors using three replicates per assay (intra-assay precision/repeatability) (ii) using two plates in parallel (intermediate precision/plate-to-plate variability) and (iii) by performing the assays on three different days (inter-assay precision/reproducibility). In addition, we determined specificity, linearity and quantification limits of the assay. Further we tested precision across the two assay systems, IFNγ/IL2 FluoroSpot and the corresponding enzymatic single cytokine ELISpot. The validation revealed: (1) a high intra-assay precision (coefficient of variation (CV) 9.96, 8.85 and 13.05 %), intermediate precision (CV 6.48, 10.20 and 12.97 %) and reproducibility (CV 20.81 %, 12,75 % and 12.07 %) depending on the analyte and antigen used; (2) a specificity of 100 %; (3) a linearity with R (2) values from 0.93 to 0.99 depending on the analyte. The testing of the precision across the two assay systems, adduced a concordance correlation coefficient p c = 0.99 for IFNγ responses and p c = 0.93 for IL2 responses, indicating a large agreement between both assay methods. The validated primary endpoint assay, an EBV peptide pool specific IFNγ/IL2 FluoroSpot assay was found to be suitable for the detection of EBV-specific immune responses subject to the requirement of standardized assay procedure and data analysis. |
X Demographics
The data shown below were collected from the profiles of 4 X users who shared this research output. Click here to find out more about how the information was compiled.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Argentina | 1 | 25% |
| Unknown | 3 | 75% |
Demographic breakdown
| Type | Count | As % |
|---|---|---|
| Members of the public | 4 | 100% |
Mendeley readers
The data shown below were compiled from readership statistics for 65 Mendeley readers of this research output. Click here to see the associated Mendeley record.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 65 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Researcher | 21 | 32% |
| Student > Master | 8 | 12% |
| Other | 5 | 8% |
| Student > Ph. D. Student | 5 | 8% |
| Student > Bachelor | 5 | 8% |
| Other | 5 | 8% |
| Unknown | 16 | 25% |
| Readers by discipline | Count | As % |
|---|---|---|
| Immunology and Microbiology | 18 | 28% |
| Agricultural and Biological Sciences | 9 | 14% |
| Biochemistry, Genetics and Molecular Biology | 6 | 9% |
| Medicine and Dentistry | 5 | 8% |
| Pharmacology, Toxicology and Pharmaceutical Science | 5 | 8% |
| Other | 5 | 8% |
| Unknown | 17 | 26% |
Attention Score in Context
This research output has an Altmetric Attention Score of 5. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 27 July 2018.
All research outputs
#6,344,611
of 23,923,788 outputs
Outputs from Journal of Translational Medicine
#977
of 4,232 outputs
Outputs of similar age
#101,993
of 357,321 outputs
Outputs of similar age from Journal of Translational Medicine
#24
of 118 outputs
Altmetric has tracked 23,923,788 research outputs across all sources so far. This one has received more attention than most of these and is in the 73rd percentile.
So far Altmetric has tracked 4,232 research outputs from this source. They typically receive a lot more attention than average, with a mean Attention Score of 10.7. This one has done well, scoring higher than 76% of its peers.
Older research outputs will score higher simply because they've had more time to accumulate mentions. To account for age we can compare this Altmetric Attention Score to the 357,321 tracked outputs that were published within six weeks on either side of this one in any source. This one has gotten more attention than average, scoring higher than 71% of its contemporaries.
We're also able to compare this research output to 118 others from the same source and published within six weeks on either side of this one. This one has done well, scoring higher than 80% of its contemporaries.