Fukui H, Matsumoto M, Bode C, Bode J C, Tsujita S, Tsujii T
Third Department of Internal Medicine, Nara Medical University, Kashihara, Japan.
J Gastroenterol Hepatol. 1993 Nov-Dec;8(6):577-81. doi: 10.1111/j.1440-1746.1993.tb01656.x.
A recently developed chromogenic endotoxin assay with plasma Tween 80 pretreatment was compared with the conventional dilution and heating method. Plasma endotoxin was measured in patients with liver cirrhosis and upper gastrointestinal (GI) bleeding by these methods. Plasma endotoxin concentration was calculated from an individual internal standard curve by adding three different standard endotoxin solutions to each sample. By the conventional heating method, added standard endotoxin gave different OD values in each sample and the slope of the standard curve showed interindividual variations. When sample plasma from chronic alcoholics was pretreated with 1% Tween 80 and ultrasonification after heating, the slope of standard curves was somewhat increased and interindividual variation was minimized. Significantly higher plasma endotoxin levels in cirrhotics with upper GI bleeding compared with those without upper GI bleeding was detected by this Tween 80 method. There was a strongly positive correlation between the endotoxin levels determined by this method and those determined by the perchloric acid method and endotoxin-specific substrate in patients with upper GI bleeding. Endotoxin levels, which were elevated 1-2 days after the bleeding, tended to decrease as patients recovered. In summary, the recovery of endogenous and exogenous endotoxin from plasma sample was increased by adding Tween 80 before the chromogenic substrate assay. Transient elevation of plasma endotoxin was demonstrated by this Tween 80 method in patients with liver cirrhosis and upper GI bleeding.
将最近开发的经血浆吐温80预处理的显色内毒素测定法与传统的稀释加热法进行了比较。采用这些方法对肝硬化合并上消化道(GI)出血患者的血浆内毒素进行了检测。通过向每个样本中添加三种不同浓度的标准内毒素溶液,根据个体内标曲线计算血浆内毒素浓度。采用传统加热法时,添加的标准内毒素在每个样本中给出的OD值不同,标准曲线的斜率显示个体间存在差异。当慢性酒精中毒患者的样本血浆在加热后用1%吐温80预处理并超声处理时,标准曲线的斜率有所增加,个体间差异最小化。采用这种吐温80法检测到,合并上消化道出血的肝硬化患者血浆内毒素水平显著高于未合并上消化道出血的患者。在合并上消化道出血的患者中,用该方法测定的内毒素水平与用高氯酸法和内毒素特异性底物测定的内毒素水平之间存在强正相关。出血后1 - 2天升高的内毒素水平,随着患者康复有下降趋势。总之,在显色底物测定前添加吐温80可提高血浆样本中内源性和外源性内毒素的回收率。采用这种吐温80法证实了肝硬化合并上消化道出血患者血浆内毒素的短暂升高。