鸡肌肉组织中左氧氟沙星残留ELISA检测方法的研究

Establishment of Indirect Competitive ELISA Method for the Determination of Levofloxacin Residues in Chicken Muscle

用N-羟基琥珀酰亚胺法把左氧氟沙星（LVL）与牛血清白蛋白（BSA）偶联制备免疫抗原,免疫新西兰大白兔得到LVL的多克隆抗体,建立了LVL间接竞争ELISA方法。结果表明：抗LVL血清效价达1∶212以上,得到标准曲线的线性回归方程为y=-0.289 4x＋0.049 5（R2=0.987 8）,50%抑制浓度（IC50）为64 ng/mL,最低检测限（LOD）为10 ng/mL,标准曲线的线性范围为10～1 000 ng/mL。批内变异系数为3.51%～7.46%,批间变异系数为8.66%～11.89%,鸡肌肉中的LVL添加浓度在10～1 000 ng/ml范围内时,回收率为73.5%～84.36%。本试验建立的ELISA方法能够满足左氧氟沙星兽药残留检测要求。

Levofloxacin（LVL） was coupled with bovine serum album（BSA） to prepare the immune antigen（LVL-BSA）by N-hydroxysuccinimide ester（NHS） method.LVL-BSA as immunizing antigen immune New-zeland albino rabbit to obtain polyclonal antibody for anti-levofloxacin.We have established indirect compete ELISA method. The result indicated that specificity antibody titer reached above 1∶212,the regression equation was y=-0.204 4x＋0.961 0（R2 = 0.985 6）,the 50% inhibition concentration（IC50） was 64 ng/mL,the limits of detect was 10 ng/mL,and the linear range about the standard curve was 10～1 000 ng/mL.The coefficients of variation for the Ci-ELISA were 3.51%～7.46% in intra-assay and 8.66%～11.89% in inter-assay.The recoveries of ofloxacin in chicken ranged from 73.5%～84.36%.This method can satisfy the residue determination of ofloxacin in chicken.