Tsuji K, Martin P A, Gaunnac G L
Prog Clin Biol Res. 1987;231:443-57.
Methods to detect endotoxin in human plasma for LAL testing have been developed by use of a perchloric acid (PCA) or a trifluoroacetic acid (TFA) treatment. These acid oxidative treatments eliminate interferences present in plasma. The mode of action of the acids for the elimination of the interferences is indicated to be by their oxidative actions. The treated plasma is assayed by use of an automated microtiter plate-chromogenic substrate LAL assay method employing a modified Cetus Pro/Pette EXPRESS system. The automated system assays 96 samples in approximately 22 minutes. Assay sensitivity of the automated Pro/Pette system is approximately 0.01 endotoxin units (EU) per ml with a linear range from 0.03 to 0.14 EU/ml. The relative standard deviation (RSD) of the assay system is approximately 6%. The improved precision of the LAL assay by the automated system has been attributed to its fast speed (2 minutes) in dispensing the unstable LAL reagent to all 96 samples in one assay block. The total operation, including the acid oxidative treatments and the 96 sample LAL assay, takes approximately 45 minutes. The recovery of spiked endotoxin from plasma ranged from 75 to approximately 100% by adding 0.4 ml of either 0.1-0.2 N PCA or 0.2-0.35 N TFA to 0.2 ml plasma and reacting for 15 minutes at 37 degrees C. The standard deviation of the assay is approximately 0.1 EU/ml plasma. The method has been proven to be rugged, simple, rapid, and cost effective thus would be suitable for clinical application.
已开发出通过使用高氯酸（PCA）或三氟乙酸（TFA）处理来检测用于鲎试剂检测的人血浆中内毒素的方法。这些酸氧化处理消除了血浆中存在的干扰物。酸消除干扰物的作用方式表明是通过它们的氧化作用。经处理的血浆通过使用采用改良的Cetus Pro/Pette EXPRESS系统的自动微量滴定板 - 显色底物鲎试剂检测方法进行检测。该自动系统在约22分钟内可检测96个样品。自动Pro/Pette系统的检测灵敏度约为每毫升0.01内毒素单位（EU），线性范围为0.03至0.14 EU/ml。该检测系统的相对标准偏差（RSD）约为6％。自动系统对鲎试剂检测精度的提高归因于其在一个检测模块中向所有96个样品分配不稳定的鲎试剂的速度快（2分钟）。整个操作，包括酸氧化处理和96个样品的鲎试剂检测，大约需要45分钟。通过向0.2 ml血浆中加入0.4 ml 0.1 - 0.2 N PCA或0.2 - 0.35 N TFA并在37℃下反应15分钟，血浆中加标内毒素的回收率在75％至约100％之间。检测的标准偏差约为每毫升血浆0.1 EU。该方法已被证明是耐用、简单、快速且具有成本效益的，因此适用于临床应用。
