Reproducibility end Correlations of Multiplex Cytokine Levels in Asymptomatic Persons
Reproducibility end Correlations of Multiplex Cytokine Levels in Asymptomatic Persons
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DOI：
10.1158/1055-9965.epi-08-0311
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发表时间：
2008-12-01
影响因子：
3.8
通讯作者：
Rabkin, Charles S.
中科院分区：
文献类型：
--
作者：
Wong, Hui-Lee;Pfeiffer, Ruth M.;Rabkin, Charles S.
基本原理：细胞因子是体液调节分子，在发病机制的免疫途径中共同起作用。某些疾病的特征是血药浓度严重升高;生理范围内的变化也可能具有重要意义。因此，我们评估了多重细胞因子immunoassay.Methods的性能特点：我们使用了基于荧光珠（Luminex）的免疫测定试剂盒，同时测量白细胞介素（IL）1 β，IL 2，IL 4，IL 5，IL 6，IL 7，IL 8，IL 10，IL 12 p70，IL 13，IFN γ，粒细胞集落刺激因子，和肿瘤坏死因子-α。我们在三个不同的日子测试了来自38名无症状个体的相同血清等分试样，以及来自另外38名预期具有低细胞因子浓度的健康供体的血清、肝素化血浆和酸性柠檬酸盐葡萄糖血浆的匹配组。我们应用多重插补来计算低于检测限的测量值的无偏重现性估计值。结果：在13种细胞因子中，有3种（IL 1 β、IL 2、IL 5）在半数以上的血清标本中未检出。重复血清测量的变异系数范围为18%至44%，组内相关系数范围为55%至98%。只有IL 4，IL 6，ILS有统计学意义的相关性（斯皮尔曼rho，0.42-0.94）之间的血清和酸性柠檬酸盐葡萄糖或肝素plasma levels.Conclusions：个体间的差异超过这些测定的大量实验室变异，产生高的组内相关系数，尽管变异系数不令人印象深刻。血浆测量值通常不能反映血清水平，因此不可互换。Luminex细胞因子检测试剂盒体积小、单次检测成本低、具有多重检测能力，因此在流行病学研究中具有潜在的实用价值。（癌症流行病学生物标志物Prev 2008;17（12）：3450-6）
Rationale: Cytokines are humoral regulatory molecules that act together in immunologic pathways underlying pathogenesis. Grossly elevated blood levels characterize certain diseases; variations within physiologic ranges could also have significance. We therefore evaluated the performance characteristics of a multiplex cytokine immunoassay.Methods: We used a fluorescent bead-based (Luminex) immunoassay kit to simultaneously measure interleukin (IL) 1 beta, IL2, IL4, IL5, IL6, IL7, IL8, IL10, IL12p70, IL13, IFN gamma, granulocyte colony-stimulating factor, and tumor necrosis factor-alpha. We tested identical aliquots of serum from 38 asymptomatic individuals on three different days and matched sets of serum, heparinized plasma, and acid citrate dextrose plasma from an additional 38 healthy donors expected to have low cytokine concentrations. We applied multiple imputation to calculate unbiased reproducibility estimates for measurements below the limits of detection. Correlations among the cytokines were assessed by Spearman rank order coefficients and principal components analyses.Results: Of the 13 cytokines, 3 were undetectable (IL1 beta, IL2, IL5) in more than half of the serum samples. Coefficients of variation for replicate serum measurements ranged from 18% to 44%, with intraclass correlation coefficients ranging from 55% to 98%. Only IL4, IL6, and ILS had statistically significant correlations (Spearman rho, 0.42-0.94) between serum and acid citrate dextrose or heparin plasma levels.Conclusions: Interindividual differences outweigh substantial laboratory variation for these assays, yielding high intraclass correlation coefficients despite unimpressive coefficients of variation. Plasma measurements generally are not reflective of serum levels and hence are not interchangeable. With their small volume, low cost per test, and multiplex capacity, Luminex-based cytokine assays have potential utility for epidemiologic studies. (Cancer Epidemiol Biomarkers Prev 2008;17(12):3450-6)