A multi-center ring trial of allergen analysis using fluorescent multiplex array technology

Eva M. King, Stephanie Filep, Bryan Smith, Thomas Platts-Mills, Robert G. Hamilton, Detlef Schmechel, Joanne E. Sordillo, Donald Milton, Ronald van Ree, Esmeralda J.M. Krop, Dick J.J. Heederik, Nervana Metwali, Peter S. Thorne, Darryl C. Zeldin, Michelle L. Sever, Agustin Calatroni, Samuel J. Arbes, Herman E. Mitchell, Martin D. Chapman

Research output: Contribution to journalArticlepeer-review

25 Scopus citations


Background: Consistent performance of allergen assays is essential to ensure reproducibility of exposure assessments for investigations of asthma and occupational allergic disease. This study evaluated intra- and inter-laboratory reproducibility of a fluorescent multiplex array, which simultaneously measures eight indoor allergens in a single reaction well. Methods: A multi-center study was performed in nine laboratories in the US and Europe to determine the inter-laboratory variability of an 8-plex array for dust mite, cat, dog, rat, mouse and cockroach allergens. Aliquots of 151 dust extract samples were sent to participating centers and analyzed by each laboratory on three separate occasions. Agreement within and between laboratories was calculated by the concordance correlation coefficient (CCC). Results: Results were obtained for over 32,000 individual allergen measurements. Levels covered a wide range for all allergens from below the lower limit of detection (LLOD = 0.1-9.8. ng/ml) to higher than 6800. ng/ml for all allergens except Mus m 1, which was up to 1700. ng/ml. Results were reproducible within as well as between laboratories. Within laboratories, 94% of CCC were ≥ 0.90, and 80% of intra-laboratory results fell within a 10% coefficient of variance (CV%). Results between laboratories also showed highly significant positive correlations for all allergens (~ 0.95, p < 0.001). Overall means of results were comparable, and inter-laboratory CV% for all allergens except Rat n 1 ranged between 17.6% and 26.6%. Conclusion: The data indicate that performance criteria for fluorescent multiplex array technology are reproducible within and between laboratories. Multiplex technology provides standardized and consistent allergen measurements that will streamline environmental exposure assessments in allergic disease.

Original languageEnglish (US)
Pages (from-to)89-95
Number of pages7
JournalJournal of Immunological Methods
Issue number1-2
StatePublished - Jan 31 2013


  • Allergen measurement
  • Asthma
  • Immunoassay
  • Indoor air quality
  • Multiplex array
  • Occupational health

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology


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