酶联免疫吸附法检测动物源性食品中恩诺沙星残留量

ELISA Determination of Enrofloxacin

为检测动物源性食品中恩诺沙星残留量,评估基于卵黄抗体的间接竞争酶联免疫吸附法检测恩诺沙星的可行性,用活性脂法将恩诺沙星同卵清蛋白偶联制备免疫原和包被抗原并用紫外光谱进行验证。用聚乙二醇-6000提取卵黄抗体。五免之后效价达到峰值1∶32 000。用间接竞争酶联免疫吸附法确定包被原质量浓度、卵黄抗体的稀释倍数和IC_(50)分别为38 ng/m L、1∶64 000和18.207 ng/m L,回归曲线方程为y=0.891 1-0.016 5x(R^2=0.990)。结果表明,制备的抗恩诺沙星卵黄抗体为进一步建立检测动物源性食品中恩诺沙星的残留的免疫方法提供参考依据。

An indirect competitive enzyme-linked immunosorbent immunoassay(ic-ELISA)based on chicken IgY was developed and validated for the detection of enrofloxacin(ENR)residues in animal-derived food samples.The ENR immunogen and coating antigen were prepared by the active ester method and confirmed by UV spectroscopy for further detection and immunization into laying hens.IgY antibodies were extracted with PEG-6000precipitation.The titer of anti-ENR IgY attained its peak(1:32000)after the fifth booster immunization.Checkerboard titration showed that1:64000dilution of anti-ENR IgY could give OD around1.0at38ng/mL ENR-OVA coating concentration.The IC50of theELISA for ENR was18.207ng/mL and the regression curve equation was y=0.8911?0.0165x(R2=0.990).The study demonstrated that anti-ENR IgY antibody was feasible for the detection of ENR in animal-derived food samples.