Variables that affect assays for plasma cytokines and soluble activation markers
Variables that affect assays for plasma cytokines and soluble activation markers
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DOI：
10.1128/cdli.6.1.89-95.1999
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发表时间：
1999-01-01
期刊：
影响因子：
--
通讯作者：
Fahey, JL
中科院分区：
文献类型：
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作者：
Aziz, N;Nishanian, P;Fahey, JL
反映体内细胞因子活性的细胞因子和可溶性免疫激活标志物越来越多地在血浆、血清和其他体液中被检测到。它们提供了有用的诊断和预后信息，以及对疾病发病机制的洞察。采用人类免疫缺陷病毒(HIV)阳性和HIV阴性受试者的血清和血浆标本，对新喋呤、β2-微球蛋白、可溶性白介素2受体、可溶性肿瘤坏死因子受体II型以及细胞因子肿瘤坏死因子α和干扰素进行检测。许多因素被发现影响这些检测的结果。当使用不同制造商的酶联免疫吸附分析(ELISA)试剂盒时，经常发现分析物的表观水平有很大差异。在某些情况下，不同制造商提供的标准存在差异。此外，由单一厂家提供的不同批次的ELISA试剂盒的分析结果差异高达50%。定量分析标准的一致性的必要性得到了清楚的说明。有细胞因子的国际参考标准，但没有可溶性细胞因子受体或可溶性激活标志物的国际参考标准。除干扰素-γ外，血清和血浆标志物水平相似。大多数分析物在几种储存条件下都是稳定的。因此，对冷冻储存的样品进行批量检测是可行的。结果表明，对于纵向研究，应使用一家制造商的预先验证的试剂来测量血浆或血清中细胞因子和免疫激活标志物的水平。实验室表现的质量可能会对临床相关性产生影响。能力测试和外部质量保证计划有助于达成所需的共识。
Cytokines and soluble immune activation markers that reflect cytokine activities in vivo are increasingly being measured in plasma, serum, and other body fluids. They provide useful diagnostic and prognostic information as well as insight into disease pathogenesis. Assays of neopterin, beta 2-microglobulin, soluble interleukin-2 receptor, and soluble tumor necrosis factor receptor type II as well as of the cytokines tumor necrosis factor alpha and gamma interferon (IFN-gamma) were evaluated by using serum and plasma samples of human immunodeficiency virus (HIV)-positive and HIV-negative subjects. Many factors were found to influence the outcomes of these assays. Substantial differences in apparent levels of analytes were frequently found when enzyme-linked immunosorbent assay (ELISA) kits from different manufacturers were used. In some cases, differences were found in the standards provided by separate manufacturers. Furthermore, the analytic results from different lots of ELISA kits supplied by single manufacturers differed by as much as 50%. The need for uniformity in the standards for quantitative assays was clearly illustrated. International reference standards are available for cytokines but not for soluble cytokine receptors or soluble activation markers. Marker levels in serum or in plasma were similar except those for IFN-gamma. Most of the analytes were stable under several storage conditions. Thus, batch testing of frozen stored samples is feasible. The findings indicate that for longitudinal studies, the levels of cytokines and immune activation markers in plasma or serum should be measured by using preverified reagents from one manufacturer. The quality of laboratory performance can have an impact on clinical relevance. Proficiency testing and external quality assurance programs can help to develop the needed consensus.