采用FIDISTM技术检测类风湿因子的多重免疫分析方法。

Multiplexed immunoassay for detection of rheumatoid factors by FIDISTM technology.

作者信息

Abreu I, Laroche P, Bastos A, Issert V, Cruz M, Nero P, Fonseca J E, Branco J, Machado Caetano J A

机构信息

Faculty of Medical Sciences, Department of Immunology, New University of Lisbon, Lisbon, Portugal.

出版信息

Ann N Y Acad Sci. 2005 Jun;1050:357-63. doi: 10.1196/annals.1313.038.

DOI:10.1196/annals.1313.038

PMID:16014552

Abstract

Our objective was to (i) compare FIDIS Rheuma, a new multiplexed immunoassay designed for simultaneous detection of IgM class rheumatoid factors (RF) directed against Fc determinants of IgG from humans and animals, with agglutination and ELISA (conventional methods) and (ii) evaluate the clinical sensitivity and specificity of biological markers for rheumatoid arthritis (RA). To do this, FIDIS technology was employed using the Luminex system. It consists of distinct color-coded microsphere sets, a flow cytometer, and digital signal processing hardware and software. Agglutination and ELISA tests were performed with commercial kits. The study included 134 samples from RA patients and 105 from healthy blood donors. For human specificity, we compared FIDIS with latex agglutination and ELISA. Relative sensitivities were 98.9% and 88.5% and specificities were 90.2% and 94.6%, respectively. For animal specificity, we compared FIDIS with Waaler-Rose and ELISA. The results were 84.9% and 71.9% for the sensitivities and 97.5% and 98.4% for the specificities, respectively. Detection of IgG anti-CCP by ELISA and IgG antikeratin by immunofluorescence was also determined in order to compare their clinical sensitivity and specificity with IgM-RF, according to the method used. The results were: IgG anti-CCP 72.3%, 97.2%; IgG antikeratin 36.6%, 100%; latex agglutination 66.4%, 97.2%; Waaler-Rose 55.9%, 96.3%; FIDIS human 73.9%, 92.1%; FIDIS animal 49.2%, 97.2%; ELISA human 93.2%, 95.5%; and ELISA animal 74.6%, 91.3%. The results showed the efficiency of FIDIS with analytical performance equivalent to the conventional methods, but having the advantage of giving quantitative results (IU/mL).

摘要

我们的目标是

（i）将一种新的用于同时检测针对人和动物IgG的Fc决定簇的IgM类类风湿因子（RF）的多重免疫分析方法FIDIS Rheuma与凝集法和酶联免疫吸附测定法（传统方法）进行比较；（ii）评估类风湿性关节炎（RA）生物标志物的临床敏感性和特异性。为此，使用Luminex系统采用了FIDIS技术。它由不同颜色编码的微球组、流式细胞仪以及数字信号处理硬件和软件组成。凝集试验和酶联免疫吸附测定法使用商用试剂盒进行。该研究纳入了134份来自RA患者的样本和105份来自健康献血者的样本。对于人类特异性，我们将FIDIS与乳胶凝集法和酶联免疫吸附测定法进行了比较。相对敏感性分别为98.9%和88.5%，特异性分别为90.2%和94.6%。对于动物特异性，我们将FIDIS与瓦勒 - 罗斯试验和酶联免疫吸附测定法进行了比较。敏感性结果分别为84.9%和71.9%，特异性结果分别为97.5%和98.4%。还根据所使用的方法通过酶联免疫吸附测定法检测IgG抗环瓜氨酸肽以及通过免疫荧光法检测IgG抗角蛋白，以便将它们的临床敏感性和特异性与IgM - RF进行比较。结果如下：IgG抗环瓜氨酸肽为72.3%，97.2%；IgG抗角蛋白为36.6%，100%；乳胶凝集法为66.4%，97.2%；瓦勒 - 罗斯试验为55.9%，96.3%；FIDIS人源为73.9%，92.1%；FIDIS动物源为49.2%，97.2%；酶联免疫吸附测定法（人源）为93.2%，95.5%；酶联免疫吸附测定法（动物源）为74.6%，91.3%。结果表明FIDIS有效，其分析性能与传统方法相当，但具有给出定量结果（IU/mL）的优势。