QUANTITATIVE LIMULUS LYSATE ASSAY FOR ENDOTOXIN AND THE EFFECT OF PLASMA
QUANTITATIVE LIMULUS LYSATE ASSAY FOR ENDOTOXIN AND THE EFFECT OF PLASMA
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DOI：
10.1136/jcp.44.10.849
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发表时间：
1991-10-01
影响因子：
3.4
通讯作者：
LOUIS, WJ
中科院分区：
文献类型：
--
作者：
HURLEY, JC;TOSOLINI, FA;LOUIS, WJ
测定了血浆和显色底物对内毒素激活鲎细胞裂解物（LAL）测定动力学的影响。在LAL试剂的浊度发展速度（光密度405）和内毒素浓度之间观察到线性相关关系，其范围超过4对数10倍。像显色底物一样，在反应中加入稀释剂和热处理过的等离子体导致光密度与存在的等离子体浓度成正比的增加。处理过的血浆的存在也导致光密度的加速增加，在测试不同浓度的血浆和血清时，结果可比较。在单个时间点监测反应进展的测定中可能无法识别这种光密度的加速增加，并且可能混淆使用显色底物的血浆样品测定。从内毒素敏感和耐药菌株的小鼠中获得的血浆显示出类似的效果。动力学方法的使用意味着可以实现血浆中内毒素的定量分析，其可变性可与半定量系列稀释相媲美，但具有更大的LAL试剂经济性。
The effects of plasma and chromogenic substrate on the kinetics of the endotoxin-activated Limulus amoebocyte lysate (LAL) assay were determined. A linear correlation was observed between the rate of development of turbidity (optical density 405) with the LAL reagent and the concentration of endotoxin over a four log ten-fold range. Like chromogenic substrate, the addition of dilution and heat treated plasma to the reaction resulted in an increase in optical density proportional to the concentration of plasma present. The presence of the treated plasma also resulted in an accelerated increase in optical density with comparable results when testing plasma at different concentrations and, additionally, serum. This accelerated increase in optical density may not be recognised in assays that monitor the progress of the reaction at a single time point and may confound assays of plasma samples that use chromogenic substrate. Plasma obtained from endotoxin sensitive and resistant strains of mice showed similar effects.The use of kinetic methodology means that a quantitative assay for endotoxin in plasma can be achieved, its variability comparable with that seen with semiquantitative serial dilution but with greater economy of the LAL reagent.