Detection of endotoxin in human blood and demonstration of an inhibitor.
Detection of endotoxin in human blood and demonstration of an inhibitor.
复制标题
人体血液中内毒素的检测和抑制剂的演示。
DOI：
--
复制
发表时间：
1970
期刊：
影响因子：
--
通讯作者：
Ronald S. Oser
中科院分区：
文献类型：
--
作者：
J. Levin;J. Levin;P. Tomasulo;P. Tomasulo;Ronald S. Oser;Ronald S. Oser
摘要鲎变形细胞裂解物的凝胶化是多种内毒素存在的敏感指标。溶解液是稳定的。溶出液与内毒素反应重现性好，易于测定。溶解液与内毒素的反应速率取决于内毒素的浓度。该反应能够检测人血浆和血清中每毫升低至0.0005 μg的内毒素。将内毒素添加到正常血液中，当将这些血液样本制备的未稀释血浆或血清与生理盐水对照进行比较时，会导致可检测到的内毒素活性丧失。凝血导致内毒素活性进一步丧失。内毒素与血液或低钙离子浓度的预孵育并不需要证明抑制作用。将血清或血浆稀释至1%至10%，几乎可以完全消除这种抑制作用。用氯仿提取血浆60分钟，基本上可以完全回收添加到全血或血浆中的内毒素。在稀释或氯仿提取血清或血浆后，阳性试验增加，表明血液的抑制性质主要是内毒素与血清蛋白之间可逆结合的结果，而不是内毒素被破坏的结果。本试验是目前所报道的体外检测内毒素最灵敏的方法。该技术在内毒素和内毒素血症的临床和实验研究中均有应用价值。
Abstract Gelation of a lysate of amebocytes of Limulus, the horseshoe crab, is a sensitive indicator of the presence of a variety of endotoxins. Amebocyte lysate is stable. The reaction between lysate and endotoxin is reproducible and easy to measure. The rate of the reaction between amebocyte lysate and endotoxin is dependent upon the concentration of endotoxin. The reaction is capable of detecting as little as 0.0005 μg per milliliter of endotoxin in human blood plasma and serum. Addition of endotoxin to normal blood resulted in loss of detectable endotoxin activity when undiluted plasma or serum prepared from such blood samples was compared with saline controls. Coagulation resulted in additional loss of endotoxin activity. Preincubation of endotoxin with blood or a low calcium ion concentration was not necessary for the demonstration of inhibition. The inhibitory effect could be eliminated almost completely by dilution of serum or plasma to 1 to 10 per cent. Extraction of plasma with chloroform for 60 minutes permitted essentially total recovery of endotoxin which had been added to whole blood or plasma. The increase in positive tests, following dilution or chloroform extraction of serum or plasma, suggested that the inhibitory nature of blood is primarily the result of reversible binding between endotoxin and serum protein(s), rather than destruction of endotoxin. This test is the most sensitive in vitro assay for endotoxin which has been reported. The technique should have application both for clinical and experimental studies of endotoxin and endotoxemia.