抗着丝粒抗体在系统性硬化症患者大队列中的研究：免疫荧光法、CENP-A 和 CENP-B ELISA 的比较。

Anti-centromere antibodies in a large cohort of systemic sclerosis patients: comparison between immunofluorescence, CENP-A and CENP-B ELISA.

机构信息

Dr. Fooke Laboratorien, Neuss, Germany.

出版信息

Clin Chim Acta. 2011 Oct 9;412(21-22):1937-43. doi: 10.1016/j.cca.2011.06.041. Epub 2011 Jul 5.

DOI:10.1016/j.cca.2011.06.041

PMID:21756890

Abstract

INTRODUCTION

Anti-centromere antibodies (ACA) are useful biomarkers in the diagnosis of systemic sclerosis (SSc) where they are found in 20-40% of patients and, albeit with lower prevalence, in patients with other systemic autoimmune rheumatic diseases. Historically, ACA were detected by indirect immunofluorescence (IIF) on HEp-2 cells and confirmed by immunoassays using recombinant CENP-B. During the last few years, to accommodate high throughput diagnostics, a number of laboratories changed from IIF to ELISA assays. The objective of this study was to compare the detection of ACA by IIF to CENP-A and a CENP-B ELISA in a large cohort of SSc patients.

METHODS

Sera collected from SSc patients (n=834) were tested for ACA by IIF on HEp-2 cells (ImmunoConcepts, Sacramento, CA) and CENP-A and CENP-B ELISA (both Dr. Fooke Laboratorien GmbH, Neuss, Germany). Furthermore, other autoantibodies were determined by QUANTA-Plex(TM) SLE 8 profile (INOVA, San Diego, CA), QUANTA Lite RNA Pol III (INOVA) and PM1-Alpha ELISA (Dr. Fooke).

RESULTS

The prevalence of ACA was 35.0% by IIF, 41.6% by CENP-A and 57.8% by CENP-B ELISA. When the CENP-A and the CENP-B ELISA results were compared to the IIF, the area under the curve value derived from receiver operating characteristic analysis was 0.98 for both assays. ACA and anti-topoisomerase I antibodies co-occurred in 1.2% (ACA by IIF), in 3.5% (by CENP-A ELISA) and in 7.4% (by CENP-B ELISA). Anti-CENP-A antibodies were negatively associated with anti-Scl-70, anti-RNA Pol III, (both p<0.0001), anti-U1-RNP (p=0.008) and anti-PM1-Alpha antibodies (p=0.0337). The degree of association was dependent on the cut-off value used.

CONCLUSION

Although we found good agreement between IIF and ELISA for the detection of ACA in SSc, a significant portion of CENP ELISA positive sera did not show the typical ACA staining pattern. Based on these findings, we conclude that an IIF ACA negative result might not rule out the presence of ACA. In addition, new CENP ELISA kits are reliable for the detection of anti-CENP in SSc sera.

摘要

简介

抗着丝粒抗体（ACA）是系统性硬化症（SSc）诊断中的有用生物标志物，在 20-40%的患者中发现，尽管在其他系统性自身免疫性风湿病患者中发现的频率较低。历史上，ACA 通过在 HEp-2 细胞上进行间接免疫荧光（IIF）检测，并使用重组 CENP-B 进行免疫测定来确认。在过去的几年中，为了适应高通量诊断，许多实验室已从 IIF 转变为 ELISA 测定。本研究的目的是比较 IIF 与 CENP-A 和 CENP-B ELISA 在大量 SSc 患者中的 ACA 检测。

方法

从 SSc 患者（n=834）收集的血清通过在 HEp-2 细胞上进行 IIF（来自 ImmunoConcepts，加利福尼亚州萨克拉门托）和 CENP-A 和 CENP-B ELISA（均来自 Dr. Fooke Laboratorien GmbH，德国 Neuss）来检测 ACA。此外，通过 QUANTA-Plex（TM）SLE 8 谱（INOVA，加利福尼亚州圣地亚哥），QUANTA Lite RNA Pol III（INOVA）和 PM1-Alpha ELISA（Dr. Fooke）确定其他自身抗体。

结果

IIF 的 ACA 患病率为 35.0％，CENP-A 为 41.6％，CENP-B ELISA 为 57.8％。当将 CENP-A 和 CENP-B ELISA 结果与 IIF 进行比较时，来自接收器操作特征分析的曲线下面积值对于两种测定均为 0.98。ACA 和抗拓扑异构酶 I 抗体同时发生在 1.2％（通过 IIF 检测到的 ACA），在 3.5％（通过 CENP-A ELISA）和 7.4％（通过 CENP-B ELISA）。抗 CENP-A 抗体与抗 Scl-70，抗 RNA Pol III（均<0.0001），抗 U1-RNP（p=0.008）和抗 PM1-Alpha 抗体呈负相关（p=0.0337）。关联程度取决于使用的截止值。

结论

尽管我们发现 IIF 和 ELISA 在 SSc 中检测 ACA 之间具有良好的一致性，但相当一部分 CENP ELISA 阳性血清未显示典型的 ACA 染色模式。基于这些发现，我们得出结论，IIF ACA 阴性结果可能不能排除 ACA 的存在。此外，新型 CENP ELISA 试剂盒可用于检测 SSc 血清中的抗 CENP。