甲萘威ELISA方法的建立及初步应用

ESTABLISHMENT AND PRIMARY APPLICATION OF ENZYME-LINKED IMMUNOADSORBENT ASSAY FOR CARBARYL

以方阵法筛选甲萘威ELISA的抗原和抗体浓度 ,并建立了甲萘威的标准抑制曲线 ;以直接竞争性ELISA为基础 ,考察了单克隆抗体的特异性和ELISA法的各种影响因素 ;在黄瓜样品中添加定量甲萘威 ,用ELISA法和HPLC法检测其中甲萘威的含量 .结果显示 ,ELISA法的最低检测限为 4 .1ng/mL ,单克隆抗体对甲萘威的特异性较强 ;甲萘威的ELISA法受分析液中离子强度、pH值、有机溶剂和样品基质浓度的影响 ;ELISA回收率在 89.4 %～ 10 3.2 %之间 ,变异系数为 7.8%～ 10 .7% ;ELISA和HPLC两种方法分析结果无显著性差异 (P >0 .0 5 ) .本研究成功建立了甲萘威的直接竞争ELISA法 ,并将该方法初步应用于黄瓜样品中甲萘威的快速检测 .图 5表 2参

The concentrations of antigen and antibody of ELISA for carbaryl were determinated with chessboard format and the inhibition curves of carbaryl were established. Based on direct competitive ELISA, the specificity of monoclonal antibodies and the factors, which may influence the ELISA results, were tested in this research. Cucumber samples were spiked with carbaryl and the concentrations were detected with both ELISA and HPLC methods. The results showed that the detection limit of ELISA was 4.1 ng/mL; monoclonal antibody was specific to carbaryl; direct competitive ELISA may be influenced by ionic concentration, pH, organic solvent and sample matrixes in assay buffer. The recoveries of ELISA were 89.4%～103.2% and coefficients of variability were 7.8%～10.7%. There were no distinct difference between the results of ELISA and HPLC (P> 0.05). A direct competitive ELISA for carbaryl was established successfully and this method was primarily applied to detect carbaryl in cumcumber samples. Fig 5, Tab 2, Ref 6