血浆中细菌内毒素定量测定方法的研究

目的:建立定量测定血浆中细菌内毒素的实验方法。方法:以内毒素检查用水制备标准曲线,动态浊度法定量测定分别采用3种抗凝剂的抗凝血中细菌内毒素的含量及回收率;以正常人血浆制备内毒素标准曲线,以抗增液复溶鲎试剂,动态浊度法定量测定人血浆中内毒素的含量及回收率。结果:使用肝素钠抗凝的血浆溶液对内毒素测定无干扰,而其他抗凝剂肝素锂、EDTA-K₂ 等有干扰;人血浆经稀释加热处理,配合使用抗增液后制备的标准曲线与使用内毒素用水制备的标准曲线相比,可以更好地测定人血浆中细菌内毒素的含量,且具有较好的重现性;用所建立的方法测定肠癌患者血浆中内毒素的含量,其结果显著高于正常人血浆中内毒素的含量。结论:以肝素钠作抗凝剂,血浆经稀释加热处理并配合使用抗增液可定量测定血浆中细菌内毒素的含量。

Objective:To establish a method to quantify the bacterial endotoxin in plasma. Methods:Kinetic-turbidimetric assay was used to determine the concentration and recovery of endotoxin in plasma anticoagulated with three different anticoagulate reagents respectively,and a standard curve of bacterial endotoxin was created by diluting endotoxin with water.In comparison,the other standard curve was created by diluting endotoxin with plasma,amoebocyte lysate was dissolved with antienhancement reagents,and kinetic-turbidimetric assay was used to quantify the endotoxin content and recovery in plasma. Results:Anticoagulate reagents such as heparin lithium and EDTA-K₂ interfered with the bacterial endotoxin test,while heparin sodium did not.The standard curve created by diluting endotoxin with diluted and heated human plasma along with amoebocyte lasate dissolved by antienhancement reagents was better at determining bacterial endotoxin in plasma than the standard curve created by diluting endotoxin with water.The standard curve was of good reproducibility,and the endotoxin content in plasma could be quantified accurately.The endotoxin content in colorectal cancer patients was much higher than that in human plasma of healthy people. Conclusions:This kinetic-turbidimetric assay requires the establishment of a standard regression curve by diluting endotoxin with diluted and heated plasma which was anticoagulated by heparin sodium.Together with antienhancement reagents,this assay can quantify the endotoxin concentration in human plasma effectively.