恩诺沙星多克隆抗体制备及阻断ELISA检测方法的研究

PREPARATION OF POLYCLONAL ANTIBODY AND DEVELOPMENT OF CIELISA FOR RAPID DETECTION OF ENROFLOXACIN

采用混合酸酐法将恩诺沙星（ENR）分别与牛血清白蛋白（BSA）和卵清白蛋白（OVA）偶联制备免疫抗原（ENR—BSA）和包被抗原（ENR—OVA）．将免疫原免疫家兔，制备多克隆抗体（ENRpAb），应用ENRpAb研制ENR残留阻断ELISA快速检测方法，并对其灵敏度、半数抑制浓度IC50、特异性、准确度进行测定．结果阻断ELISA的线性检测范围为1～205ng／mL，灵敏度1ng／mL，半数抑制浓度（IC50）为11ng／mL，与其他化合物的交叉反应率（CR％）均〈0．1％，鸡肉样的平均添加回收率分别为87．35％，平均变异系数均〈10％．本检测方法具有快速、敏感、特异、简便等特点，适合ENR残留快速检测的推广应用．

hnmunogen and covering antigen were prepared by mixed anhydride method coupling enrofloxacin to BSA or OVA. ENR pAb were generated by immunized rabbits with BSA-ENR. A ciELISA kit for detect ENR （ENR-Kit） was developed with ENR pAb and its sensitivity, IC50 , veracity, specificity were tested respectively. The ENR-Kit had the linear detection range of 1 -205ng/ml, the sensitivity of lng/ml and a good sensitivity with an IC50 of 11 ng/ml to ENR, less than 0.01% cross-reactivity to other compounds. The recoveries of ENR spiked in chicken were 87.35%. The coefficient variation was below 10%. The ENR-Kit possesses rapidity, sensitivity, specificity and briefness that were proved to be used for the rapid detection of ENR residues in animal food.