Webster C J
J Clin Microbiol. 1980 Nov;12(5):644-50. doi: 10.1128/jcm.12.5.644-650.1980.
Some factors affecting the use of chromogenic substrates with Limulus lysate for assaying bacterial endotoxins in blood have been assessed. It was found that endogenous amidases, which degrade the substrate, could be inactivated by heating serum at 60 degrees C for 15 min. Endotoxin was found not to be removed from serum during clotting. A potent inhibitor of the activated lysate was found to be anti-thrombin II, but specific absorption of anti-thrombin II from plasma reduced only marginally the inhibition of lysate by plasma. The presence of specific antibody to the endotoxin was found not to affect its ability to activate lysate. Inactivation of endotoxin by serum enzymes was biphasic in unheated serum, and most of the activity was destroyed in 3 h at 37 degrees C or in 24 h at 5 degrees C. The relevance of these findings to the objective quantitation of endotoxin activities is discussed.
已评估了一些影响使用鲎试剂显色底物检测血液中细菌内毒素的因素。发现可通过将血清在60℃加热15分钟来灭活降解底物的内源性酰胺酶。研究发现，凝血过程中血清内毒素不会被去除。抗凝血酶II被发现是活化鲎试剂的一种强效抑制剂，但从血浆中特异性吸附抗凝血酶II仅略微降低了血浆对鲎试剂的抑制作用。内毒素特异性抗体的存在不影响其激活鲎试剂的能力。在未加热的血清中，血清酶对内毒素的灭活呈双相性，大部分活性在37℃下3小时或5℃下24小时被破坏。讨论了这些发现与内毒素活性客观定量的相关性。
