Impact of clinical sample handling and processing on ultra-low level measurements of plasma cytokines
Impact of clinical sample handling and processing on ultra-low level measurements of plasma cytokines
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DOI：
10.1016/j.clinbiochem.2019.01.001
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发表时间：
2019-03-01
影响因子：
2.8
通讯作者：
Walt, David R.
中科院分区：
文献类型：
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作者：
Cohen, Limor;Keegan, Alissa;Walt, David R.
目的：在这项研究中，我们评估了临床样本的处理和处理对IL-6，IL-10，IFN γ和IL-2测量plasma.Design和Methods的影响：我们收集了全血样本，并分析了各种预分析参数。我们评估了以下内容：1）在室温和4 ℃下储存10小时的全血中的细胞因子稳定性; 2）血浆中的细胞因子稳定性超过6小时; 3）剧烈的样品处理，包括重复滴加和通过气动输送系统输送;和4）血浆中细胞因子的冻融稳定性。为了确保能够测量IL-6，IL-10，IFN γ和IL-2水平的血浆中，我们使用Simoa，一个超灵敏的免疫测定platform.Results：我们表明，全血储存在室温下的结果在降低细胞因子水平和全血储存在4摄氏度的结果在更大的细胞因子稳定性。我们还表明，细胞因子是稳定的，当全血样品进行有力的样品处理。最后，我们表明，细胞因子在血浆中是稳定的超过三个冻融cycle.Conclusions：临床样本处理和加工可以影响测量IL-6，IL-10，IFN γ，IL-2在血浆中。我们相信，这项研究将是一个有用的参考未来的研究中，这些细胞因子被用作潜在的生物标志物。
Objectives: In this study, we evaluated the impact of clinical sample handling and processing on IL-6, IL-10, IFN gamma, and IL-2 measurements in plasma.Design and methods: We collected whole blood samples and analyzed various pre-analytical parameters. We assessed the following: 1) cytokine stability in whole blood that was stored over a ten-hour period at room temperature and 4 degrees C; 2) cytokine stability in plasma over 6 h; 3) vigorous sample handling including repeated dropping and transport through a pneumatic transport system; and 4) freeze-thaw stability of cytokines in plasma. To ensure ability to measure IL-6, IL-10, IFN gamma, and IL-2 levels in plasma, we used Simoa, an ultra-sensitive immunoassay platform.Results: We show that whole blood storage at room temperature results in decreased cytokine levels and that whole blood storage at 4 degrees C results in greater cytokine stability. We also show that cytokines are stable when whole blood samples are subjected to vigorous sample handling. Lastly, we show that cytokines are stable in plasma over three freeze-thaw cycles.Conclusions: Clinical sample handling and processing can affect measurements of IL-6, IL-10, IFN gamma, and IL-2 in plasma. We believe this study will be a useful reference for future studies in which these cytokines are used as potential biomarkers.