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Comparative Study
. 2005 Aug;66(2):175-91.
doi: 10.1016/j.jri.2005.03.005.

Validation and comparison of luminex multiplex cytokine analysis kits with ELISA: determinations of a panel of nine cytokines in clinical sample culture supernatants

Nefertiti C dupont  1 , Kehui Wang, Pathik D Wadhwa, Jennifer F Culhane, Edward L Nelson
Affiliations
Comparative Study

Validation and comparison of luminex multiplex cytokine analysis kits with ELISA: determinations of a panel of nine cytokines in clinical sample culture supernatants

Nefertiti C dupont et al. J Reprod Immunol. 2005 Aug.

Abstract

Problem: Analyses of the expression pattern of multiple cytokines are frequently required for characterization of the status of the immune system as it pertains to Th type bias and intrinsic levels of inflammation. Classically, analysis of cytokine expression patterns has been performed by enzyme-linked immunosorbent assays (ELISA) for each separate analyte. A new technology, Luminex MAP, facilitates the simultaneous evaluation of multiple immune mediators with advantages of higher throughput, smaller sample volume, and lower cost. Validation of this technology has been limited to small sample sets, limited use of clinical study specimens, and use of non-commercial reagents.

Methods: Ninety-six specimens from women over the course of their respective pregnancies were evaluated for cytokine concentrations using commercially available ELISA kits and commercially available Luminex MAP kits according to the manufacturers' directions. Correlations between data sets were evaluated using Pearson's correlation coefficient (r).

Results: Excellent correlations were demonstrated for IL-1 beta, IL-4, IL-5, IL-6, IL-10, IFN gamma, and TNF alpha, in contrast to IL-12 p 70 and IL-13.

Conclusions: Luminex multiplex technology has distinct advantages and is a valid alternative method to ELISA for the evaluation of the majority of cytokines tested and for the characterization of immune system status.

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Figures

Fig. 1
Fig. 1
Scatter plots of ELISA and LINCOplex cytokine kit IL-1β, IL-4, IL-5, IL-6, IL-10 and IL-13 determinations. Panel A represents data obtained for IL-1β, r = 0.8380. Panel B represents data obtained for IL-4, r = 0.8686. Panel C, represents data obtained for IL-5, r = 0.8117. Panel D, represents data obtained for IL-6, r = 0.9029. Panel E represents data obtained for IL-10, r = 0.8204. Panel F represents data obtained for IL-13, r = 0.6193. Best-fit trend lines are depicted for each graph. All samples (n = 96) are included in these analyses.
Fig. 2
Fig. 2
Scatter plots of ELISA and multiplex cytokine kit Th1-associated cytokine determinations. Panel A represents data obtained for IFN γ, r = 0.9388. Panel B, represents data obtained for TNF α, r = 0.9377. Panel C represents data obtained for the LINCOplex kit determination of IL-12 p70, r = 0.002. Panel D represents data obtained for the Beadlyte kit determination of IL-12 p70, r = 0.8570. Best-fit trend lines are depicted for graphs of INF γ, TNF α, and Beadlyte IL-12 p70. All samples (n = 96 for Linco, n = 48 for Beadlyte) are included in these analyses.
Fig. 3
Fig. 3
Scatter plots of ELISA and Beadlyte multiplex cytokine kit determinations. Panel A represents data obtained for IL-1β, r = 0.9347. Panel B represents data obtained for IL-4, r = 0.9409. Panel C represents data obtained for IL-10, r = 0.8815. Panel D represents data obtained for IL-13, r = 0.8599. Panel E represents data obtained for IFN γ, r = 0.9256. Panel F represents data obtained for TNF α, r = 0.9510. Best-fit trend lines are depicted for each graph. All samples (n = 48) are included in these analyses.
Fig. 4
Fig. 4
Scatter plots of LINCOplex cytokine and Beadlyte multiplex cytokine kit determinations. Panel A represents data obtained for IL-1β, r = 0.9824. Panel B represents data obtained for IL-4, r = 0.9147. Panel C represents data obtained for IL-10, r = 0.9410. Panel D represents data obtained for IL-13, r = 0.9565. Panel E represents data obtained for IFN γ, r = 0.9825. Panel F represents data obtained for TNF α, r = 0.9057. Best-fit trend lines are depicted for each graph. All samples analyzed with both kits (n = 48) are included in these analyses.
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