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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https://hdl.handle.net/10161/14683Published Version (Please cite this version)
10.1016/j.jim.2014.04.011Publication Info
Lynch, Heather E; Sanchez, Ana M; D'Souza, M Patricia; Rountree, Wes; Denny, Thomas
N; Kalos, Michael; & Sempowski, Gregory D (2014). Development and implementation of a proficiency testing program for Luminex bead-based
cytokine assays. Journal of Immunological Methods, 409. pp. 62-71. 10.1016/j.jim.2014.04.011. Retrieved from https://hdl.handle.net/10161/14683.This is constructed from limited available data and may be imprecise. To cite this
article, please review & use the official citation provided by the journal.
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Show full item recordScholars@Duke
Thomas Norton Denny
Professor in Medicine
Thomas N. Denny, MSc, M.Phil, is the Chief Operating Officer of the Duke Human Vaccine
Institute (DHVI), Associate Dean for Duke Research and Discovery @RTP, and a Professor
of Medicine in the Department of Medicine at Duke University Medical Center. He is
also an Affiliate Member of the Duke Global Health Institute. Previously, he served
on the Health Sector Advisory Council of the Duke University Fuquay School of Business.
Prior to joining Duke, he was an Associate Professor of Pathology, Labo
Gregory David Sempowski
Professor in Medicine
Dr. Sempowski earned his PhD in Immunology from the University of Rochester and was
specifically trained in the areas of inflammation, wound healing, and host response
(innate and adaptive). Dr. Sempowski contributed substantially to the field of lung
inflammation and fibrosis defining the roles of pulmonary fibroblast heterogeneity
and CD40/CD40L signaling in regulating normal and pathogenic lung inflammation. During
his postdoctoral training with Dr. Barton F. H
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