阿特拉津高灵敏酶联免疫吸附检测法的建立

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（HPLC）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。

High sensitive ELISA for detection of atrazion

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.