鸡肌肉组织中左氧氟沙星兽药残留ELISA检测方法的研究

Establishment of Indirect Competitive Enzyme-linked Immunosorbent Assay for Levofloxacin in Chicken Muscle Tissue

关键词:  左氧氟沙星  多克隆抗体  间接竞争ELISA  鸡肌肉组织

Key words:  Key words Levofloxacin  Polyclonal antibody  indect Key words Levofloxacin  Polyclonal antibody  indect competitive ELISA  chicken muscle tissue

摘要:

摘要：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血清效价达达1：212以上，得到标准曲线的线性回归方程为Y = -0.2894X+0.0495(R2 = 0.9878)，50%抑制浓度（IC50）为64ng/mL，最低检测限（LOD）为10ng/mL，标准曲线的线性范围为10~1000ng/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10~1000ng/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。

Abstract:

Abstract Levofloxacin(LVL) was coupled with carrier protein 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 to reach the above of 1：212.The regression equation, the 50% inhibition concentration(IC50), the limits of detect (LOD) and the linear range about the standard curve of Ofloxacin were y=-0.2044x+0.9610 (R2 = 0.9856), 64ng/ml, 10ng/ml and 10~1000ng/ml, respectively. 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 were detected , the recoveries in chicken ranged from 73.5% to 84.36%.So our method will satisfy the determination of residues of ofloxacin in Chicken.