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用于杀虫剂杀螟硫磷的酶联免疫吸附测定。半抗原构象和样品基质对测定性能的影响。

Enzyme-linked immunosorbent assays for the insecticide fenitrothion. Influence of hapten conformation and sample matrix on assay performance.

作者信息

Kim Yoo Jung, Kim Young Ah, Lee Yong Tae, Lee Hye-Sung

机构信息

Department of Food Science and Nutrition, Kyungpook National University, Daegu 702-701, Republic of Korea.

出版信息

Anal Chim Acta. 2007 May 22;591(2):183-90. doi: 10.1016/j.aca.2007.03.072. Epub 2007 Apr 7.

Abstract

This study aimed at developing competitive enzyme-linked immunosorbent assays (ELISAs) for the organophosphorus (OP) insecticide fenitrothion using a monoclonal antibody. The hapten used to obtain the antibody had an ideal structural feature that allowed minimal functional group sacrifice. By using the antibody and a coating antigen, a competitive indirect ELISA was developed, which showed an IC50 of 14 ng mL(-1) with a detection limit of 3.0 ng mL(-1). A competitive direct ELISA using an enzyme tracer was also developed, which showed an IC50 of 17 ng mL(-1) with a detection limit of 1.6 ng mL(-1). The antibodies in both assays showed negligible cross-reactivity with the metabolites of fenitrothion and other OP pesticides except with the insecticides parathion-methyl and parathion-ethyl. Recoveries of fenitrothion from fortified rice and lettuce samples were determined and the bias in the recovery values was rationalized by using the standard curves obtained in the matrix extract.

摘要

本研究旨在利用单克隆抗体开发用于有机磷杀虫剂杀螟硫磷的竞争性酶联免疫吸附测定法(ELISA)。用于获得抗体的半抗原具有理想的结构特征,可使官能团牺牲最小化。通过使用该抗体和包被抗原,开发了一种竞争性间接ELISA,其IC50为14 ng mL(-1),检测限为3.0 ng mL(-1)。还开发了一种使用酶标记物的竞争性直接ELISA,其IC50为17 ng mL(-1),检测限为1.6 ng mL(-1)。两种测定法中的抗体与杀螟硫磷的代谢物和其他有机磷农药的交叉反应可忽略不计,但与甲基对硫磷和乙基对硫磷杀虫剂除外。测定了强化大米和生菜样品中杀螟硫磷的回收率,并通过使用基质提取物中获得的标准曲线使回收率值的偏差合理化。

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