【摘要】 制备出3种不同结合比的酶标抗体,对检测条件进行了优化,建立了标记抗体固相抗原直接竞争ELISA(cdELISA)检测丁草胺的方法。结果表明,酶标抗体克分子比为1.55时偶合物活性最高,包被浓度1μg·mL-1,酶标抗体800倍稀释,稀释液采用2×PBS,不含BSA、甲醇,建立的标准曲线IC50=1.7ng·mL-1,检测限为0.006ng·mL-1,检测范围0.06～2616.7ng·mL-1,批内变系数9.25%,批间变异系数23.55%。蒸馏水以及稀释10倍后的矿泉水、池塘水、稻田水添加1ng·mL-1和10ng·mL-1的丁草胺,用所建立的方法检测,平均回收率分别为93.27%、158.50%、119.75%、124.51%。本研究为进一步开发丁草胺免疫分析试剂盒奠定了基础。更多还原

【Abstract】 A competitive enzyme-linked immunosobent assay(ELISA)for the detection of the herbicide butachlor was development,which based on enzyme-labeled antibody and solid antigen(cdELISA).Three molar ratio of enzyme-labeled antibodies were prepared,the one with 1.55 of molar ratio had higher activity,the optimized cdELISA performed with coating antigen of 1 μg·mL-1 and enzyme conjugate of 800-fold dilution in 2×PBS dilution solution without protecting agent BSA and solvent methanol,has a butachlor IC50 of 1.7 ng·mL-1 and detection limit of 0.006 ng·mL-1,the detection range was 0.06～2 616.7 ng·mL-1,the inter CV was 9.25% and intra CV was 23.55%.The water samples after 10-fold dilution,fortified with butachlor,were analyzed according to this mothod.The average recovery rate for mineral water,pond water,rice filed water and distilled water was 158.50%、119.75%、124.51%、93.27%,respectively,which proved the suitability of the method to develop commercial immunoassay kit for butachlor.更多还原