Validation of assays for inflammatory mediators in exhaled breath condensate
Validation of assays for inflammatory mediators in exhaled breath condensate
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DOI：
10.1183/09031936.00081707
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发表时间：
2008-05-01
影响因子：
24.3
通讯作者：
Stockley, R. A.
中科院分区：
文献类型：
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作者：
Bayley, D. L.;Abulsiriwil, H.;Stockley, R. A.
使用呼出气冷凝液（EBC）作为非侵入性评估肺部炎症的工具正变得越来越普遍。许多作者使用商业ELISA试剂盒来测量EBC内的炎症介质。然而，EBC中非常低浓度的介质通常低于相关ELISA的商业验证浓度范围，并且关键地低于S形标准曲线的线性部分。本研究旨在验证一系列用于EBC的检测方法，并比较EBC与相应的溶胶相痰标本的检测结果，用ELISA法检测了以下介质：白三烯（LT）B-4、白细胞介素（IL）-8、分泌性白细胞蛋白酶抑制剂和α（1）-抗胰蛋白酶（AAT）。仅19.6%的受试者在一种测定法（AAT）中介质浓度达到定量下限，而在三种测定法（LTB 4、IL-8和AAT分别为31、6.5和61%）中介质浓度达到检测下限。没有显着的相关性之间存在任何介质在EBC和溶胶阶段sputter. Results本研究表明，护理时，必须行使解释介质测量呼出气冷凝液和化验必须验证在相关的浓度内发现的生物流体。
The use of exhaled breath condensate (EBC) as a tool for noninvasive assessment of lung inflammation is becoming commonplace. Many authors use commercial ELISA kits to measure inflammatory mediators within EBC. However, the very low concentrations of mediators within EBC are often below the commercially validated concentration range of the relevant ELISA and crucially below the linear part of the sigmoid standard curve. The present study seeks to validate a series of assays for use in EBC and to compare the results in EBC with those from matched sol phase sputum samples.The following mediators were measured by ELISA: leukotriene (LT)B-4, interleukin (IL)-8, secretory leukoprotease inhibitor and alpha(1)-antitrypsin (AAT). Myeloperoxidase was measured by chromogenic substrate assay.Mediator concentrations reached the lower limit of quantification in only one assay (AAT) in 19.6% of subjects, while mediator concentrations reached the lower limit of detection in three assays (LTB4, IL-8 and AAT in 31, 6.5 and 61% of subjects, respectively). No significant correlations were present between any mediators in EBC and sol phase sputum.The results of the present study indicate that care must be exercised when interpreting mediator measurements in exhaled breath condensate and that assays must be validated at concentrations relevant to those found within the biological fluid.