Background/objective

Cytokines and chemokines (cytokines) are central to rheumatoid arthritis (RA) pathogenesis, with increasing use of multiplex immunoassays in clinical/research settings. Rheumatoid factor (RF) may interfere with assay outcomes by nonspecifically binding detection analytes. We evaluated the performance of a commercially available multiplex platform, including assessment of the impact of RF depletion.

Methods

Forty-five cytokines were tested using Meso Scale Discovery V-PLEX™ and samples from 40 RA and 40 osteoarthritis (OA) patients. Select samples were depleted of RF using a commercial binder. Performance was assessed using intra-assay coefficients of variation (CV), intraclass correlation coefficients (ICC), percent change following RF depletion, and disease discrimination. Values above or below quantification thresholds were imputed.

Results

Of the 45 cytokines analyzed, 31 yielded CVs <10%; none demonstrated CVs >30%. ICCs universally exceeded 0.85 with the exception of eight analytes. RF depletion altered cytokine values by <15% for 40 analytes with larger changes (>30%) only seen for one analyte. Twenty-three cytokines differed significantly based on measurement in plasma vs. serum. Three analytes were higher in the serum of RA vs. OA (IL-10, IP-10, TNFα), and none were significantly greater in OA vs. RA. Seventeen analytes required imputation for >50% of the samples tested, primarily related to concentrations below the lower limit of quantification threshold.

Conclusion

The results from this commercially available multiplex assay were generally highly reproducible and interference induced by RF only meaningfully impacted the quantification of five of the analytes examined.

中文翻译：

---

在评估类风湿性关节炎和骨关节炎患者循环细胞因子和趋化因子中的商业化多重平台的性能

背景/目标

细胞因子和趋化因子（细胞因子）是类风湿性关节炎 (RA) 发病机制的核心，在临床/研究环境中越来越多地使用多重免疫测定。类风湿因子 (RF) 可能会干扰非特异性结合检测分析物的检测结果。我们评估了商用多路复用平台的性能，包括评估射频损耗的影响。

方法

使用 Meso Scale Discovery V-PLEX™ 和来自 40 名 RA 和 40 名骨关节炎 (OA) 患者的样本测试了 45 种细胞因子。使用商业粘合剂去除精选样品的 RF。使用分析内变异系数 (CV)、组内相关系数 (ICC)、RF 耗尽后的百分比变化和疾病鉴别来评估性能。估算高于或低于量化阈值的值。

结果

在分析的 45 种细胞因子中，31 种产生的 CV <10%；没有一个显示 CV > 30%。除八种分析物外，ICC 普遍超过 0.85。RF 消耗使 40 种分析物的细胞因子值改变了 <15%，只有一种分析物的变化较大 (>30%)。23 种细胞因子在血浆与血清中的测量值显着不同。RA 与 OA 血清中的三种分析物（IL-10、IP-10、TNFα）较高，而 OA 与 RA 中没有一种分析物显着较高。17 种分析物需要对 >50% 的测试样品进行插补，主要与低于量化阈值下限的浓度有关。

结论

这种商业上可用的多重测定的结果通常具有高度可重复性，并且由 RF 引起的干扰仅对所检查的五种分析物的定量产生有意义的影响。