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

Journal Article (Journal Article;Review)

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.

Full Text

Duke Authors

Cited Authors

  • Lynch, HE; Sanchez, AM; D'Souza, MP; Rountree, W; Denny, TN; Kalos, M; Sempowski, GD

Published Date

  • July 2014

Published In

Volume / Issue

  • 409 /

Start / End Page

  • 62 - 71

PubMed ID

  • 24801479

Pubmed Central ID

  • PMC4138252

Electronic International Standard Serial Number (EISSN)

  • 1872-7905

Digital Object Identifier (DOI)

  • 10.1016/j.jim.2014.04.011


  • eng

Conference Location

  • Netherlands