Development and implementation of a proficiency testing program for Luminex bead-based cytokine assays.
Abstract
Luminex bead array assays are widely used for rapid biomarker quantification due to the ability to measure up to 100 unique analytes in a single well of a 96-well plate. There has been, however, no comprehensive analysis of variables impacting assay performance, nor development of a standardized proficiency testing program for laboratories performing these assays. To meet this need, the NIH/NIAID and the Cancer Immunotherapy Consortium of the Cancer Research Institute collaborated to develop and implement a Luminex assay proficiency testing program as part of the NIH/NIAID-sponsored External Quality Assurance Program Oversight Laboratory (EQAPOL) at Duke University. The program currently monitors 25 domestic and international sites with two external proficiency panels per year. Each panel includes a de-identified commercial Luminex assay kit with standards to quantify human IFNγ, TNFα, IL-6, IL-10 and IL-2, and a series of recombinant cytokine-spiked human serum samples. All aspects of panel development, testing and shipping are performed under GCLP by EQAPOL support teams. Following development testing, a comprehensive site proficiency scoring system comprised of timeliness, protocol adherence, accuracy and precision was implemented. The overall mean proficiency score across three rounds of testing has remained stable (EP3: 76%, EP4: 75%, EP5: 77%); however, a more detailed analysis of site reported results indicates a significant improvement of intra- (within) and inter- (between) site variation, suggesting that training and remediation for poor performing sites may be having a positive impact on proficiency. Through continued proficiency testing, identification of variables affecting Luminex assay outcomes will strengthen efforts to bring standardization to the field.
Type
Journal ArticleSubject
CytokinesGCLP
Luminex
Multiplex
Proficiency
Biomarkers
Cooperative Behavior
Cytokines
Guideline Adherence
High-Throughput Screening Assays
Humans
International Cooperation
Laboratories
Laboratory Proficiency Testing
Monitoring, Immunologic
Multicenter Studies as Topic
Observer Variation
Practice Guidelines as Topic
Predictive Value of Tests
Program Development
Program Evaluation
Quality Control
Quality Improvement
Quality Indicators, Health Care
Reference Standards
Reproducibility of Results
Specimen Handling
Time Factors
Workflow
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http://hdl.handle.net/10161/14683Published Version (Please cite this version)
10.1016/j.jim.2014.04.011Collections
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