甲拌磷残留检测间接竞争ELISA试剂盒的研制-食品科学.PDF

284 2011, Vol. 32, No. 04 食品科学 ※技术应用 魏松红，逄若霖，王 翀，纪明山，谷祖敏，祁之秋，王英姿 (沈阳农业大学植物保护学院，辽宁 沈阳 110866) 研制用于检测甲拌磷残留量的间接竞争 ELISA 试剂盒。甲拌磷包被抗原的最佳工作质量浓度为 4mg/L ，抗 体的最佳稀释倍数为 8000，甲拌磷多克隆抗体交叉反应率小于20% ，甲拌磷抑制率回归曲线为y =18.846x ＋6.9949， 在 1～5000 μg/L 范围内呈线性关系，检测限为4.90 μg/L ，IC50 为 191.37μg/L 。甲拌磷试剂盒的添加回收率均高于 75% ，供试样品检测结果的批内和批间变异系数均低于 10%。试剂盒在4 ℃或－20 ℃下有效期为270d 。 甲拌磷；残留；酶联免疫吸附测定法；试剂盒 Development of Indirect ELISA Kit for Detecting Phorate Residue WEI Song-hong ，PANG Ruo-lin ，WANG Chong ，JI Ming-shan ，GU Zu-min ， QI Zhi-qiu ，WANG Ying-zi (College of Plant Protection, Shenyang Agricultural University, Shenyang 110866, China) Abstract：An indirect enzyme-linked immunosorbent assay(ELISA) kit was developed to monitor phorate residue. The optimal concentration of coating antigen was 4 mg/L and the optimal dilution ratio of antibody was 8000. The cross-reaction ratio was less than 20%. The regression equation of inhibition rate for phorate was y = 18.846x ＋6.9949 with a linear detection range of 1 －5000 μg/L. Results indicated that the detection limit was 4.90 μg/L and IC50 was 191.37 μg/L. The recovery rates were higher than 75%. The intra-assay and inter-assay coefficients were less than 10%. The developed kit can be stored at 4 ℃ or －20 ℃ for 270 days. Key words ：phorate ；residue ；enzyme-linked immunosorbent assay(ELISA)；kit 中图分类号：S482.4 文献标识码：B 文章编号：1002-6630(2011)04-0284-04 甲拌磷[phorate，O,O- 二乙基 -S-( 乙硫基甲基)二硫 异性，IC50 为 0.12～3.8 μg/mL 。本实验在甲拌磷自主合 代磷酸酯] 是有机磷类重要的内吸性杀虫杀螨剂，主要 成人工抗原及制备多克隆抗体的研究基础上[7] ，研制甲 用于防治棉花、水稻、小麦、高粱等大田作物上的害 拌磷间接竞争 ELISA 试剂盒，并对其性能进行测定，适 虫[ 1 ] ，为土壤和种子处理剂，高等毒性。甲拌磷在植 用于土壤、粮食等样品中甲拌磷残留的分析。 物体内氧化为更高毒性的氧化物，并有较长的残效期，