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基于单克隆抗体的茚虫威残留检测用灵敏间接竞争酶联免疫吸附测定法(ic-ELISA)的建立。

Development of a sensitive Indirect Competitive Enzyme-Linked Immunosorbent Assay (ic-ELISA) based on the monoclonal antibody for the detection of the imidaclothiz residue.

机构信息

Department of Pesticide Science, College of Plant Protection, Nanjing Agricultural University , Jiangsu Key Laboratory of Pesticide Science, Key Laboratory of Monitoring and Management of Crop Diseases and Pest Insects, Ministry of Agriculture, Nanjing 210095, People's Republic of China.

出版信息

J Agric Food Chem. 2011 Mar 9;59(5):1594-7. doi: 10.1021/jf104241n. Epub 2011 Feb 3.

Abstract

An indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) based on monoclonal antibodies (MoAbs) for imidaclothiz was developed. The hapten of imidaclothiz was synthesized and conjugated to bovine serum albumin (BSA) and ovalbumin (OVA) to form the artificial antigens. MoAbs were obtained by immunizing BALB/c mice. Under the optimized conditions (10% methanol, 0.14 M Na(+), and pH 7.4), the half-maximal inhibition concentration (IC(50)) was 0.0875 ± 0.0034 mg/L and the limit of detection (IC(20)) was 0.0178 ± 0.0018 mg/L for imidaclothiz. There were no obvious cross-reactivities with most of the structural analogues of neonicotinoid insecticides, except imidacloprid. The recoveries of imidaclothiz in environmental and agricultural samples, including tap water, paddy water, soil, and cabbage, ranged from 80.43 to 113.83%, well within the requirements of residue detection. These results showed that this immunoassay could be used for the determination of imidaclothiz in environmental and agricultural samples.

摘要

建立了一种基于单克隆抗体(MoAb)的间接竞争酶联免疫吸附法(ic-ELISA),用于检测咪鲜胺。合成了咪鲜胺的半抗原,并将其与牛血清白蛋白(BSA)和卵清蛋白(OVA)偶联,形成人工抗原。通过免疫 BALB/c 小鼠获得了 MoAb。在优化条件下(10%甲醇、0.14 M Na(+)和 pH 7.4),咪鲜胺的半抑制浓度(IC(50))为 0.0875 ± 0.0034 mg/L,检测限(IC(20))为 0.0178 ± 0.0018 mg/L。除了吡虫啉外,与大多数新烟碱类杀虫剂的结构类似物没有明显的交叉反应。咪鲜胺在环境和农业样品中的回收率,包括自来水、稻田水、土壤和白菜,范围在 80.43%至 113.83%之间,完全符合残留检测的要求。这些结果表明,该免疫分析方法可用于环境和农业样品中咪鲜胺的测定。

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