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目的建立细菌内毒素的多粘菌素B(PMB)与抗内毒素兔多克隆抗体联合夹心ELISA(sELISA)检测法。方法将PMB包被于酶标板作固相,封闭后逐次加入不同浓度的大肠杆菌内毒素,优化抗内毒素兔多抗和酶标抗体的浓度,建立sELISA方法。结果在0.1～100μg/ml范围内,内毒素浓度与OD值呈正相关关系,回归方程为y=0.094lnx+0.227,R2=0.914,最低检测限为100 ng/ml。结论该方法适用于内毒素检测。
Objective To establish a sandwich enzyme - linked immunosorbent assay (ELISA) for detection of polymyxin B (PMB) and polyclonal antibody against endotoxin in rabbits. Methods PMB was coated on a microtiter plate for solid phase. After blocking, different concentrations of E. coli endotoxin were added successively to optimize the polyclonal antibody against rabbit polyclonal antibody against endotoxin and the concentration of enzyme-labeled antibody. The sELISA method was established. Results In the range of 0.1 ~ 100μg / ml, the concentration of endotoxin was positively correlated with OD. The regression equation was y = 0.094lnx + 0.227, R2 = 0.914, and the detection limit was 100 ng / ml. Conclusion This method is suitable for endotoxin testing.