Multiplex bead array assays for detection of soluble cytokines: Comparisons of sensitivity and quantitative values among kits from multiple manufacturers

Authors

  • Sameena S. Khan,

    1. Warren G. Magnuson Clinical Center, Critical Care Medicine Department, National Institutes of Health, Bethesda, Maryland
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  • Meghan S. Smith,

    1. National Heart, Lung, and Blood Institute, Flow Cytometry Core Facility, National Institutes of Health, Bethesda, Maryland
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  • Debra Reda,

    1. Warren G. Magnuson Clinical Center, Critical Care Medicine Department, National Institutes of Health, Bethesda, Maryland
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  • Anthony F. Suffredini,

    1. Warren G. Magnuson Clinical Center, Critical Care Medicine Department, National Institutes of Health, Bethesda, Maryland
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  • J. Philip McCoy Jr.

    Corresponding author
    1. National Heart, Lung, and Blood Institute, Flow Cytometry Core Facility, National Institutes of Health, Bethesda, Maryland
    • National Institutes of Health, National Heart, Lung, and Blood Institute, Flow Cytometry Core, Building 10, Room 4A07, Bethesda, MD 20892
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  • This paper is a contribution of the U.S. National Institutes of Health and is not subject to copyright. Certain commercial equipment, instruments, materials, or companies are identified in this paper to specify the experimental procedure. Such identification does not imply recommendation or endorsement by National Institutes of Health, nor does it imply that the material or equipment identified are the best available for this purpose.

  • This article is a US government work and, as such, is in the public domain in the United States of America.

Abstract

Background

Multiplex bead array assays permit simultaneous cytometric quantitation of multiple cytokines in solution by capturing these to spectrally distinct beads. Because several manufacturers offer reagents to quantitate the same cytokines on a single instrument, a comparison should be made to determine whether these kits yield similar data and whether these data are comparable to enzyme-linked immunosorbent assay (ELISA).

Methods

This study compared cytokine detection kits by using Luminex 100. Twenty-six serum samples from seven subjects were analyzed for interferon-γ, interleukins 1β, 6, and 8, and tumor necrosis factor-α by using multiplex kits from LINCO Research, Bio-Rad Laboratories, R&D Systems, and BioSource International. Each assay was performed according to the manufacturers' specifications. Standard curves were generated by using reference concentrations supplied by each manufacturer. ELISAs for interleukin-8 were performed by using kits from R&D and BioSource.

Results

Cytokine levels followed similar patterns, although absolute concentrations differed among kits. ELISA and Luminex values for interleukin-8 were similar in kits from the same manufacturer.

Conclusions

Because relative cytokine measurements are often valuable when performed serially, it may be possible to make interlaboratory comparisons by using different kits. When comparison of absolute values is crucial, kits from the same supplier should be used. Within-vendor, bead array, and ELISA values appear comparable. Published 2004 Wiley-Liss, Inc.

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