内毒素的多粘菌素B与抗体联用夹心ELISA检测法  

Detection of Endotoxin by sELISA Using Conjunction of Polymyxin B and Polyclonal Antibody against Endotoxin

作　　者：周卓晟[1,2] 罗中捷[1,3] 郝春慧[1] 郭寅生[1] 刘红艳[1] 黄正[1] 

机构地区：[1]华中科技大学同济医学院公共卫生学院环境与健康教育部重点实验室,湖北武汉430030 [2]深圳市龙岗区卫生监督分所 [3]广州市卫生监督所

出　　处：《环境与健康杂志》2012年第2期185-186,共2页Journal of Environment and Health

基　　金：国家自然科学基金(30771774)

摘　　要：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。Objectives Sandwich ELISA （sELISA） using polymyxin B （PMB） and anti-endotoxin polyclonal antibody was built to detect endotoxin. Methods Polymyxin B was coated onto ELISA plates. After blocking the different concentrations of endotoxin, polyclonal antibody against endotoxin and HRP-labelled antibody were added into each well to establish the sELISA. Results The consequence of sELISA was as follow： the concentration of endotoxin from 0.1 μg/ml to 100 μg/ml was in direct proportion to the value of OD and showed a linear regression equation： y=0.094 lnx＋0.227, R2 was 0.914 and the detection limit was 100 ng/ml. Conclusion The method was suitable for detecting endotoxin.

关 键 词：内毒素 多粘菌素B 多克隆抗体 sELISA 

分 类 号：R117[医药卫生—公共卫生与预防医学]