管式磁微粒化学发光免疫分析法测定玉米样品中的黄曲霉毒素B1

Development of a sensitive magnetic particles chemiluminescence enzyme immunoassay for determination of aflatoxin B1 in corn

开发了一种管式磁微粒化学发光免疫分析法测定玉米样品中黄曲霉毒素B1的方法,该方法使待测玉米样品中的黄曲霉毒素B1、辣根过氧化物酶标记的黄曲霉毒素B1与异硫氰酸荧光素(FITC)标记的黄曲霉毒素B1单克隆抗体在均相体系中发生竞争性免疫反应,再加入用抗FITC抗体包被的磁微粒作分离剂,抗原抗体复合物结合在磁微粒上,在磁场中经分离、洗涤后加发光底物,检测发光强度,测定玉米样品中黄曲霉毒素B1的含量.此方法标准曲线线性范围为0.05～5ng/mL,检测限为0.02ng/mL,批内相对标准偏差小于9%,批间相对标准偏差小于15%,具有良好的稳定性和重现性.

A sensitive magnetic particles chemiluminescence enzyme immunoassay for determination of aflatoxin B1（AFB1） in corn was developed and validated.The method was developed based on a competitive immunoreaction format.Immunocomplex was formed through the competitive immunoreaction among AFB1 antigen,horseradish peroxidase-labeled AFB1 antigen and fluorescein isothiocyanate labeled anti-AFB1 monoclonal antibody.Immunocomplex combined with anti-fluorescein isothiocyanate （FITC） antibody coated magnetic particles.The magnetic particles served as both the solid phase and separator.The horseradish peroxidase（HRP）-luminol-hydrogen peroxide chemiluminescent system was chosen as the detection system.This method had a detection limit of 0.02 ng/mL and a linear range of 0.05-5 ng/mL.The intra-assay relative standard deviations were less than 9%.The interassay relative standard deviations were less than 15%.The recoveries were from 94.7% to 109%.The method has been successfully applied to the detection of aflatoxin B1（AFB1） in corn.