Journal
CLINICA CHIMICA ACTA
Volume 339, Issue 1-2, Pages 147-156Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.cccn.2003.10.002
Keywords
fluorescence immunochromatography assay; laser-fluorescence scanner; BCG dye-binding method; human serum albumin
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Background: Human serum albumin (HSA) is the most abundant plasma protein and plays key a role in metabolism. The variation in albumin concentration provides valuable information related to metabolic diseases and diagnostic application. Methods: We constructed two assay systems to quantify the albumin concentration. The immunoassay used a fluorescence (FL) dye to detect albumin in samples and employed the conventional chromatography as a separation system. The assay system consists of an anti-HSA-mAb or an HSA immobilized test strip in a disposable cartridge, a fluorescence-labeled detector buffer and a laser-fluorescence scanner. We mixed the sample with detector, loaded it onto a cartridge, incubated it for 10 min and measured the concentration of albumin in a laser-fluorescence scanner. We examined the comparability of assay with an automated BCG dye binding method using a Hitachi 747 biochemical analyzer. Results: The correlation of coefficient between A(T)/A(C) as converted from the relative fluorescence units (RFU) and albumin concentration displayed reasonable reliability in both the competition and the inhibition assay systems (r = 0.998). Using the Bland-Altman difference plot analysis, we observed an acceptable agreement between two methods, the fluorescence immunochromatography assay (FL-ICA) and the automated BCG dye-binding method of a Hitachi biochemical analyzer, over the clinical relevant range of HSA concentrations. The coefficient of variation (CV) of within- and between-run variation in the immunoassay system was <8% and the recovery fell within 5% in each control sample. In addition to its reliable analytical performance, the assay with whole blood can be completed in 12 min using a one-step operation without any pretreatment. Conclusion: The developed immunoassay system using fluorescence dye and lateral-flow chromatography is a simple, fast and reliable method for quantifying the albumin concentration in whole blood. (C) 2003 Elsevier B.V. All rights reserved.
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