摘要
采用自制的抗伏马毒素B1(FB1)多克隆抗体,建立定量检测玉米中FB1的化学发光间接竞争ELISA方法。通过化学偶联分别获得免疫抗原钥孔血蓝蛋白(KLH)-FB1和包被抗原卵清蛋白(OVA)-FB1。用KLH-FB1免疫新西兰大白兔,制备多克隆抗体。以OVA-FB1为包被抗原,FB1为竞争抗原,建立检测FB1的间接竞争化学发光ELISA方法,并对该方法进行了条件优化。该方法对FB1标品进行检测,得到回归方程为Y=0.602 5-0.370 7 X,相关系数R2为0.986 2,检测限为0.14ng/mL,线性关系良好的检测范围为0.14～33.3ng/mL,玉米样品加标回收率83.5%～102%。表明建立的化学发光ELISA法敏感性好、特异性高,能用于玉米中FB1毒素的检测。
To develop an indirect competitive chemiluminescence enzyme linked immunosorbent assay （CL- ELISA） for the detection of fumonisin B1 （FB1） in corn samples, conjugate antigens with keyhole limpet hemocyanin （KLH）-FB1 and ovalbumin （OVA）-FB1 were synthesized, polyclonal antibody was prepared by immunizing the New Zealand rabbits with KLH-FB1, and OVA-FB1 was used as coating antigen. After optimizing the working conditions and factors, CL-ELISA was established and used to detect FB1 in spiked corn sample with FB1 standard substance. The results showed that, using the CL-ELISA method to determine FB1, the regression equation was Y=0. 602 5--0. 370 7X （ R2= O. 986 2）, the detectable range was from 0. 14 ng/mL to 33. 3 ng/mL with a minimum detection limit of 0. 14 ng/mL, and the recovery reached 83.5%-102%. It can be concluded that the method we developed is sensitive and hopefully can be used to the actual determination of FB1 in corn samples.
出处
《上海交通大学学报（农业科学版）》
2012年第5期45-50,共6页
Journal of Shanghai Jiaotong University(Agricultural Science)
基金
国家863计划项目(2007AA10Z424)
