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Clinical Chemistry and Laboratory Medicine (CCLM)

Published in Association with the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM)

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Analytical performance of a multiplexed, bead-based cytokine detection system in small volume samples

Martin Bomert1, a / Gabi Köllisch1, a / Marjut Roponen2 / Roger Lauener3, 4 / Harald Renz1 / Petra I. Pfefferle1 / 1

1Institute of Laboratory Medicine and Pathobiochemistry, Molecular Diagnostics, Philipps University, Marburg, Germany

2Department of Environmental Health, National Institute for Health and Welfare, Kuopio, Finland

3University of Zürich, Children’s Hospital, and Christine Kühne-Center for Allergy Research and Education, Zürich, Switzerland

4Children’s Allergy and Asthma Hospital, Hochgebirgsklinik, and Christine Kühne-Center for Allergy Research and Education, Davos, Switzerland

aMartin Bomert and Gabi Köllisch contributed equally to this work.

Corresponding author: PD Dr. Nadia Al-Fakhri, Institute of Laboratory Medicine and Pathobiochemistry, Molecular Diagnostics, Philipps University Marburg, Baldingerstr., 35043 Marburg, Germany Phone: +49-6421-5866265, Fax: +49-6421-5866189

Citation Information: Clinical Chemistry and Laboratory Medicine. Volume 49, Issue 10, Pages 1691–1693, ISSN (Online) 1437-4331, ISSN (Print) 1434-6621, DOI: https://doi.org/10.1515/CCLM.2011.631, June 2011

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Background: Multiplexed cytokine measurement offers many advantages over the conventional enzyme-linked immunosorbent assay (ELISA) format when applied in large- scale epidemiological studies or clinical trials. In the present study we set out to define the reliability and consistency of a suspension multiplexed protein array, the cytometric bead array (CBA), in large-scale, longitudinal studies.

Methods: The cytokines interleukin (IL)-5, IL-10, tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ) were measured in pediatric samples from childhood asthma and allergy studies. Analytical performance of CBA was determined in sample supernatants and CBA was compared to conventional ELISA.

Results: Within-run and total imprecision were between 5.2%–10.8% and 5.6%–13.2%, respectively, at three different concentrations for all cytokines. Slopes of dilution linearity were between 1.01 and 1.31 for the four cytokines. The recovery rate at two different concentrations of the cytokines was between 97% and 113%. Lower limits of detection and quantification as well as functional sensitivity were determined. Comparison of the multiplex array and solid phase method showed good correlation with r between 0.82 and 0.93. The sample volume required for the multiplex format was 25% of the ELISA sample volume.

Conclusions: CBA analytical evaluation and comparison to an ELISA format demonstrated high reproducibility, sensitivity and good applicability for small volume samples.

Keywords: allergy; analytical performance; cytokine measurement; immunology; multiplex assay system

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