Multiplex assay reliability and long-term intra-individual variation of serologic inflammatory biomarkers

Heather S. McKay*, Joseph B. Margolick, Otoniel Martínez-Maza, Joseph Lopez, John Phair, Giovanna Rappocciolo, Thomas N. Denny, Larry I. Magpantay, Lisa P. Jacobson, Jay H. Bream

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

34 Scopus citations


Background Circulating cytokines, chemokines, and soluble cytokine receptors can serve as biomarkers of inflammation and immune dysregulation. Good reliability of multiplex platforms, which allow for simultaneous, comprehensive biomarker assessment, is critical for their utility in epidemiologic studies. We examined the reliability of the Meso-Scale Discovery (MSD) platform to simultaneously quantitate 15 cytokines and chemokines and the Luminex platform (R&D Systems) to quantitate 5 soluble receptors and 2 chemokines and cytokines and evaluated long-term within-person correlation of these biomarkers. Methods The detectability and reliability of these assay systems were assessed using the same external controls across plates and archived sera from 250 HIV men in the Multicenter AIDS Cohort Study. Using up to four visits per person from 1984 to 2009, age-adjusted intraclass correlation coefficients (ICC) of biomarkers with >80% detectability (CCL11, CXCL8, CXCL10, CCL2, CCL4, CCL13, CCL17, CXCL13, IL-10, IL-12p70, IL-6, TNF-α, BAFF, sCD14, sCD27, sgp130, sIL-2Rα, and sTNF-R2) were obtained using linear mixed models. Results Most biomarkers were detectable in 80% of control samples; IFN-γ, GM-CSF, and IL-2 were undetectable in >20% of samples. Among the HIV-uninfected men, most biomarkers showed fair to strong within-person correlation (ICC > 0.40) up to 15 years. The ICC for CXCL8 was good in the short term but decreased with increasing time between visits, becoming lower (ICC < 0.40) after 8 years. Conclusions These multiplexed assays showed acceptable reliability for use in epidemiologic research, despite some technical variability and limitations in cytokine quantitation. Most biomarkers displayed moderate-to-excellent intra-individual variability over the long term, suggesting their utility in prospective studies investigating etiologic associations with diverse chronic conditions.

Original languageEnglish (US)
Pages (from-to)185-192
Number of pages8
StatePublished - Feb 1 2017


  • Inflammatory biomarkers
  • Intraclass correlation
  • Multiplex assay reliability
  • Reliability

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology
  • Biochemistry
  • Hematology
  • Molecular Biology


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