恩诺沙星ELISA快速检测方法的建立  

To Establish the ELISA Method for Detecting Enrofloxacin Residues

作　　者：刘红[1] 曾振灵[2] 杨桂香[2] 陈杖榴[2] 

机构地区：[1]中山市农产品质量监督检验检测中心,广东中山528403 [2]华南农业大学兽医学院,广东广州510642

出　　处：《中国兽药杂志》2006年第11期13-15,共3页Chinese Journal of Veterinary Drug

基　　金：广东省自然科学基金资助项目(994162)

摘　　要：采用活酯法合成酶标记半抗原（ENR-HRP），紫外扫描分析和ELISA方法鉴定，结合比为1．6：1。选择ENR-HRP与游离ENR相竞争的模式建立检测ENR残留的ELISA检测方法，其最佳工作条件为：二抗的包被浓度为2μg／mL，抗体工作浓度1：1000，酶标半抗原工作浓度1：500。标准曲线在0．423～1000ng／mL之间线性关系良好，其IC50为33．76ng／mL，回归方程为Y=-0．07769lnx＋0．770801，相关系数r=0．99153，最低检测限0．423ng／mL。单抗对达氟沙星、培氟沙星、麻保沙星有少许交叉，特异性较好。ELISA检测方法批内平均变异系数为5．75％，批间平均变异系数为10．69％，重复性较好。ENR-HRP conjugation was synthesized by ester activation method. The synthesized reactant （ENR- HRP） was analyzed by UV scan and detected by ELISA. The ratio of ENR to HRP of ENR-HRP was 1.6： 1. A competitive ELISA was developed to detect the residues of enrofloxacin by competing of ENR-HRP and free ENR. The optimal working conditions for ELISA were as follows：the concentration of the goat anti-mouse lgG （for coating） was 2 μg/mL, the titer of monoclonal antibody was 1 ： 1 000 ; the titer of ENR-HRP was 1 ： 500. The standard curve had a good linearity in the range of 0.423 ～ 1 000 ng/mL. The IC50 was 33.76 ng/mL, and the detection limit was 0. 423 ng/mL. The regression equation was y = -0. 077 69ln x ＋0. 770 801, and the correlation coeffi- cient was 0.991 53. And the antibody had a little cross-reaction against danofloxacin, pefloxacin and marbofloxacin. The intra-assay variation of the ELISA method was 5.75%, and the inter-assay variation was 10.69%, which indicated a good repetition for the ELISA method.

关 键 词：恩诺沙星 酶标记半抗原 单克隆抗体 ELISA 

分 类 号：S859.84[农业科学—临床兽医学]