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Clinical Chemistry and Laboratory Medicine (CCLM)

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

Editor-in-Chief: Plebani, Mario

Ed. by Gillery, Philippe / Lackner, Karl J. / Lippi, Giuseppe / Melichar, Bohuslav / Payne, Deborah A. / Schlattmann, Peter / Tate, Jillian R.

12 Issues per year


IMPACT FACTOR 2016: 3.432

CiteScore 2016: 2.21

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1437-4331
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Volume 49, Issue 10 (Oct 2011)

Issues

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

Martin Bomert
  • Institute of Laboratory Medicine and Pathobiochemistry, Molecular Diagnostics, Philipps University, Marburg, Germany
  • Martin Bomert and Gabi Köllisch contributed equally to this work.
  • Other articles by this author:
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/ Gabi Köllisch
  • Institute of Laboratory Medicine and Pathobiochemistry, Molecular Diagnostics, Philipps University, Marburg, Germany
  • Martin Bomert and Gabi Köllisch contributed equally to this work.
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  • De Gruyter OnlineGoogle Scholar
/ Marjut Roponen
  • Department of Environmental Health, National Institute for Health and Welfare, Kuopio, Finland
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/ Roger Lauener
  • University of Zürich, Children’s Hospital, and Christine Kühne-Center for Allergy Research and Education, Zürich, Switzerland
  • Children’s Allergy and Asthma Hospital, Hochgebirgsklinik, and Christine Kühne-Center for Allergy Research and Education, Davos, Switzerland
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/ Harald Renz
  • Institute of Laboratory Medicine and Pathobiochemistry, Molecular Diagnostics, Philipps University, Marburg, Germany
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/ Petra I. Pfefferle
  • Institute of Laboratory Medicine and Pathobiochemistry, Molecular Diagnostics, Philipps University, Marburg, Germany
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/ Nadia Al-Fakhri
  • Institute of Laboratory Medicine and Pathobiochemistry, Molecular Diagnostics, Philipps University, Marburg, Germany
  • Email
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Published Online: 2011-06-11 | DOI: https://doi.org/10.1515/CCLM.2011.631

Abstract

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

About the article

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


Received: 2011-03-08

Accepted: 2011-05-05

Published Online: 2011-06-11

Published in Print: 2011-10-01


Citation Information: Clinical Chemistry and Laboratory Medicine, ISSN (Online) 1437-4331, ISSN (Print) 1434-6621, DOI: https://doi.org/10.1515/CCLM.2011.631.

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