使用荧光免疫测定法（I）对全血中的人白蛋白进行快速、简单的测量。

A rapid, simple measurement of human albumin in whole blood using a fluorescence immunoassay (I).

作者信息

Choi Sunga, Choi Eui Yul, Kim Dong Joon, Kim Jae Hoon, Kim Tai Sun, Oh Sang Wook

机构信息

Central Research Institute of BodiTech Inc., Chuncheon 200-957, South Korea.

出版信息

Clin Chim Acta. 2004 Jan;339(1-2):147-56. doi: 10.1016/j.cccn.2003.10.002.

DOI:10.1016/j.cccn.2003.10.002

PMID:14687905

Abstract

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 AT/AC 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.

摘要

背景

人血清白蛋白（HSA）是血浆中含量最丰富的蛋白质，在新陈代谢中起关键作用。白蛋白浓度的变化为代谢性疾病和诊断应用提供了有价值的信息。

方法

我们构建了两个检测系统来定量白蛋白浓度。免疫测定法使用荧光（FL）染料检测样品中的白蛋白，并采用传统色谱作为分离系统。该检测系统由一个固定有抗HSA单克隆抗体或HSA的一次性试剂盒中的试纸条、荧光标记的检测缓冲液和激光荧光扫描仪组成。我们将样品与检测器混合，加载到试剂盒上，孵育10分钟，然后在激光荧光扫描仪中测量白蛋白浓度。我们使用日立747生化分析仪，通过自动BCG染料结合法检验了该检测方法的可比性。

结果

在竞争和抑制检测系统中，由相对荧光单位（RFU）转换而来的AT/AC与白蛋白浓度之间的相关系数显示出合理的可靠性（r = 0.998）。使用Bland-Altman差异图分析，我们发现在HSA浓度的临床相关范围内，荧光免疫层析测定法（FL-ICA）和日立生化分析仪的自动BCG染料结合法这两种方法之间具有可接受的一致性。免疫测定系统批内和批间变异的变异系数（CV）< 8%，每个对照样品的回收率在5%以内。除了可靠的分析性能外，全血检测可通过一步操作在12分钟内完成，无需任何预处理。