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Evaluation of an automated immunoassay method for cytokine measurement using the Immulite Immunoassay system.

Abstract：

:Cytokines are key mediators in cell regulation and communication. The concentration of these proteins can rapidly and importantly increase during severe clinical situations. However, current techniques are not adapted to stat measurement, thus making their clinical use limited. In this context, the commercialization of five new kits for cytokine measurement interleukin ((IL)-1, IL-6, IL-8, tumor necrosis factor (TNF)-alpha and IL-2R) on an automated immunoanalyzer, the Immulite, seems to be a new approach for the determination of these markers. We report here the evaluation of the performance of these tests. The technique is based on a solid phase (bead) two site chemiluminescent enzyme immunometric assay. The analysis is performed within 60 to 90 minutes and the calibration is stable for 15 days. The values of the between-run imprecision study were similar to those from the within-run study with coefficients of variation (CV) ranging from 2% (low values of IL-8) to 11.5% for intermediate concentrations of IL-6 (500 pg/ml). CVs were usually around 5%. The accuracy was determined by a linearity study using standards (except for IL-2R) provided by the National Institute for Biological Standards and Control (NIBSC). Slopes obtained during this study were close to 1 (r2 = 0.99), except for IL-6, for which the slope was 1.55. TNF-alpha values were close to those expected. IL-1 results were about 20% higher. IL-6 values were over estimated above 100 pg/ml and under estimated below this value. IL-8 study seemed to be impaired by the poor stability of this molecule in the NIBSC preparation. Correlation study with standard laboratory techniques gave variable results: for IL-1 (n = 43) the slope was 0.77 (study carried out using cell culture media), for IL-6 (n = 54) the slope was 0.78, for IL-8 (n = 37) the slope was 1.64, for TNF-alpha (n = 40) the slope was 0.33 and the slope for IL-2R (n = 51) was 5.1. For the last cytokine, the unit in Immulite assay was different from the one used in our comparison technique. Cross-calibration results were consistent with these data and show that the bias is probably linked to a calibration problem. The study demonstrated excellent practicality of the system, and good stability of the calibration curve (15 days). However, the sample volume required (350 microl for the IL-6 and the TNF-alpha) could constitute a limitation for pediatric measurements.

journal_name

Clin Chem Lab Med

journal_title

Clinical chemistry and laboratory medicine

authors

Berthier F,Lambert C,Genin C,Bienvenu J

doi

10.1515/CCLM.1999.092

keywords:

subject

Has Abstract

pub_date

1999-05-01 00:00:00

pages

593-9

issue

eissn

1434-6621

issn

1437-4331

journal_volume

pub_type

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Evaluation of an automated immunoassay method for cytokine measurement using the Immulite Immunoassay system.