Evaluation of multiplexed fluorescent microsphere immunoassay for detection of autoantibodies to nuclear antigens
Evaluation of multiplexed fluorescent microsphere immunoassay for detection of autoantibodies to nuclear antigens
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DOI：
10.1128/cdli.11.6.1054-1059.2004
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发表时间：
2004-11-01
期刊：
影响因子：
--
通讯作者：
Hill, HR
中科院分区：
文献类型：
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作者：
Martins, TB;Burlingame, R;Hill, HR
可提取核抗原（ENA）的抗体存在于多种胶原血管疾病中。确定这些抗体的个体特异性在建立疾病诊断和在某些情况下的预后方面非常有用。用多重荧光微球免疫测定法，在249份血清样本中测定了对五种最具诊断价值的ENA的反应性，其中包括来自56名先前记录为系统性红斑狼疮（SLE）患者的样本。多重检测的结果与已建立的ENA酶联免疫吸附试验（ELISA）的结果进行了比较，所评价的5种ENA的一致性、灵敏度和特异性分别为：SSA，99.1、100.0和98.8%; SSB，98.69、88.9和99.5%; Sm，97.6、95.8和97.9%; RNP 97.2、92.7和98.8%; Scl-70、93.6、50.0和99.0%。在56例确诊的SLE患者中，多重检测的自身抗体显著浓度的频率为SSA 21.4%，SSB 7.1%，Sm 10.7%，RNP 32.1%，Scl-70 0%。对于本研究中研究的五种ENA标志物中的四种，新的基于流式细胞仪微珠的多重检测与成熟的单分析物ELISA方法显示出良好的相关性。两种检测方法之间最显著的差异是Scl-70抗原，其最常被解决，有利于多重检测。我们的研究表明，基于多重微球的免疫分析方法是一种敏感和特异性的检测和半定量人血清中ENA抗体的方法。
Antibodies to extractable nuclear antigens (ENA) are found in a variety of collagen vascular diseases. Determining the individual specificities of these antibodies is extremely useful in establishing the disease diagnosis and in some cases the prognosis. With a multiplexed fluorescent microsphere immunoassay, reactivity to five of the most diagnostically useful ENA was measured in 249 serum samples, including samples from 56 patients previously documented to have systemic lupus erythematosus (SLE). Results of the multiplexed assay were compared to results from established ENA enzyme-linked immunosorbent assays (ELISAs), and the agreement, sensitivity, and specificity, respectively, for the five ENA evaluated were as follows: SSA, 99.1, 100.0, and 98.8%; SSB, 98.69 88.9, and 99.5%; Sm, 97.6, 95.8, and 97.9%; RNP 97.2, 92.7, and 98.8%; Scl-70, 93.6, 50.0, and 99.0%. In the 56 confirmed SLE patients, the frequency of significant concentrations of autoantibodies with the multiplexed assay was 21.4% for SSA, 7.1% for SSB, 10.7% for Sm, 32.1% for RNP, and 0% for Scl-70. The new flow cytometric bead-based multiplexed assay showed excellent correlation with the well-established single-analyte ELISA methods for four of five the ENA markers investigated in this study. The most notable discrepancies between the two assays were for the Scl-70 antigen, which was most often resolved in favor of the multiplexed assay. Our studies show that the multiplexed microsphere-based immunoassay is a sensitive and specific method for the detection and semiquantitation of ENA antibodies in human sera.