阿特拉津高灵敏酶联免疫吸附检测法的建立

摘要

Objective:To set up ELISA for detection of atrazine with high precision.Methods:The reaction condition of indirect-ELISA was optimized,including atrazine-ovalbumin(AT-OVA) concentration and primary antibody concentration,organic solvent,goat anti-rat immunoglobin G-horseradish peroxidase(IgG-HRP) concentration.The actual samples were detected by the ELISA method established in our laboratory.Then the ELISA method was compared with the HPLC.Results:The specification curve of indirect-ELISA was set up after optimization.The relation coefficient R2 =0.9958.The limit of detection (LOD) was 1.972 ng/ml.The percent recovery of the actual samples was in range of 80％ ～ 120％.The ELISA detection sensitivity was higher than the HPLC in the range of 0 ng/ml ～ 6 ng/ml atrazine.Conclusion:The ELISA to detect atrazine has good specificity and high precision.And it can be applied in testing real atrazine samples replacing of the large-scale instrument.%目的:建立高灵敏度的阿特拉津酶联免疫吸附检测法.方法:将间接竞争ELISA进行条件优化以提高检测灵敏度,包括包被抗原与一抗的最佳工作浓度筛选、选择一抗的最佳稀释度对包被抗原进行细化筛选、不同有机溶剂对竞争结合反应的影响、酶标二抗稀释度筛选等.用建立的酶联免疫检测法检测实际样品,再与高效液相色谱法(HrLC)检测进行比较.结果:利用优化后条件建立了阿特拉津间接竞争ELISA检测曲线,标准曲线的相关系数R2=0.9958,相关性较好.另由此标准曲线可得LOD(最低检出限)为1.972 ng/ml.用于检测实际样品,回收率在80％-120％之间.当添加样品浓度为(0～6)ng/ml时,该法的检测灵敏度高于HPLC.结论:新建立的阿特拉津ELISA特异性好、精密度高,可代替大型仪器用于阿特拉津实际样品检测.