摘要
报道一种新型显色基质Boc-Leu-Gly-Arg-DEAA的合成和应用该基质建立的鲎试验法.测定游离显色基质DEAA与1-萘酚-2-磺酸缩合后形成的蓝色物质的吸收度与内毒素浓度的关系.实验表明,蓝色物质在675nm波长处的吸收度与内毒素的浓度在一定范围内呈线性关系.该法比凝胶鲎试验法的灵敏度高10倍,一般能检测0.006EU/ml的内毒素,若延长保温反应时间可检测含量低至0.002EU/ml的内毒素.测定样品的组内误差为4.8%,组间误差为6.8%.在测定病人血浆中的内毒素时不受任何病人血浆色泽的干扰,故特别适用于临床.
This paper reports the synthesis of a new chromogenic substrate Boc-Leu-Gly-ArgDEAA and introduces the finding of a new chromogenic Limulus assay for the quantitation of endotoxins using the substrate.This assay is based on the property that the synthetic chromogenic substrate can be hy drolysed by enzymes in the Limulus amebocyte lysate activated by endotoxins, releasing a chromogen DEAA which can be further coupled to 1-naphthol-2-sulfonate to form a blue-colored substance.It is show that the absorbance of the blue-colored substance at the wavelength of 675 nm is directly proportion al to the concentration of the endotoxin solution, thus enabling the quantitation of endotoxins.The chro mogenic method is ten times more sensitive than the clot-gel method, and it can generally detect as little as 0.006EU standard endotoxin per ml.The advantage of this assay is its avoidance of the interference from the plasma colors of patients.Thus, it is particularly applicable for the clinical quantitation of endo toxins.
基金
国家自然科学基金
关键词
鲎试验
显色基质
内毒素
limulus assay
chromogenic substrate
endotoxin
