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Development and implementation of a proficiency testing program for Luminex bead-based cytokine assays.

Publication ,  Journal Article
Lynch, HE; Sanchez, AM; D'Souza, MP; Rountree, W; Denny, TN; Kalos, M; Sempowski, GD
Published in: J Immunol Methods
July 2014

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.

Duke Scholars

Published In

J Immunol Methods

DOI

EISSN

1872-7905

Publication Date

July 2014

Volume

409

Start / End Page

62 / 71

Location

Netherlands

Related Subject Headings

  • Workflow
  • Time Factors
  • Specimen Handling
  • Reproducibility of Results
  • Reference Standards
  • Quality Indicators, Health Care
  • Quality Improvement
  • Quality Control
  • Program Evaluation
  • Program Development
 

Citation

APA
Chicago
ICMJE
MLA
NLM
Lynch, H. E., Sanchez, A. M., D’Souza, M. P., Rountree, W., Denny, T. N., Kalos, M., & Sempowski, G. D. (2014). Development and implementation of a proficiency testing program for Luminex bead-based cytokine assays. J Immunol Methods, 409, 62–71. https://doi.org/10.1016/j.jim.2014.04.011
Lynch, Heather E., Ana M. Sanchez, M Patricia D’Souza, Wes Rountree, Thomas N. Denny, Michael Kalos, and Gregory D. Sempowski. “Development and implementation of a proficiency testing program for Luminex bead-based cytokine assays.J Immunol Methods 409 (July 2014): 62–71. https://doi.org/10.1016/j.jim.2014.04.011.
Lynch HE, Sanchez AM, D’Souza MP, Rountree W, Denny TN, Kalos M, et al. Development and implementation of a proficiency testing program for Luminex bead-based cytokine assays. J Immunol Methods. 2014 Jul;409:62–71.
Lynch, Heather E., et al. “Development and implementation of a proficiency testing program for Luminex bead-based cytokine assays.J Immunol Methods, vol. 409, July 2014, pp. 62–71. Pubmed, doi:10.1016/j.jim.2014.04.011.
Lynch HE, Sanchez AM, D’Souza MP, Rountree W, Denny TN, Kalos M, Sempowski GD. Development and implementation of a proficiency testing program for Luminex bead-based cytokine assays. J Immunol Methods. 2014 Jul;409:62–71.
Journal cover image

Published In

J Immunol Methods

DOI

EISSN

1872-7905

Publication Date

July 2014

Volume

409

Start / End Page

62 / 71

Location

Netherlands

Related Subject Headings

  • Workflow
  • Time Factors
  • Specimen Handling
  • Reproducibility of Results
  • Reference Standards
  • Quality Indicators, Health Care
  • Quality Improvement
  • Quality Control
  • Program Evaluation
  • Program Development