化学发光酶联免疫分析法同时检测3种有机磷农药残留

Development and evaluation of chemiluminescence enzyme-linked immunoassay for residue detection of three organophosphorus pesticides

利用能同时识别对硫磷、甲基对硫磷和杀螟硫磷的宽谱特异性单克隆抗体,建立了同时测定这3种有机磷农药残留的化学发光酶联免疫分析方法 (chemiluminescence enzyme immunoassay,CLEIA),比较了间接竞争(ic-CLEIA)和直接竞争(dc-CLEIA)2种反应模式,优化了相关理化参数,确立了最适反应条件。结果表明:间接竞争CLEIA法检测对硫磷、甲基对硫磷和杀螟硫磷的抑制中浓度(IC_(50))分别为5.57、2.30和2.62μg/kg,检测线性范围分别为0.39~100、0.39~25和0.39~25μg/kg;直接竞争CLEIA法检测对硫磷、甲基对硫磷和杀螟硫磷的抑制中浓度(IC_(50))分别为5.43、1.34和1.24μg/kg,检测线性范围分别为0.39~100、0.10~25和0.10~25μg/kg。所建立的CLEIA方法基本能满足对硫磷、甲基对硫磷和杀螟硫磷在谷物和果蔬中最大残留限量的检测要求,为研制有机磷农药多残留检测试剂盒提供了技术依据。

In this study, chemiluminescence enzyme-linked immunoassay（CLEIA） for the detection of the three organophosphorus pesticides was established and optimized. A monoclonal antibody with broad-specificity to parathion, parathion-methyl, and fenitrothion was employed. The direct and indirect competitive CLEIAs（dc-CLEIA and ic-CLEIA） were compared, and the physical and chemical parameters in those two reaction models were optimized. The half-maximum inhibition concentration（IC50） and the linear range from ic-CLEIA was respectively 5.57 μg/kg and 0.39-100 μg/kg for parathion, 2.30 μg/kg and 0.39-25 μg/kg for parathion-methyl, and 2.62 μg/kg and 0.39 to 25 μg/kg for fenitrothion. The half-maximum inhibition concentration（IC50） and the linear range for dc-CLEIA was respectively 5.43 μg/kg and 0.39-100 μg/kg for parathion, 1.34 μg/kg and 0.10-25 μg/kg for parathionmethyl, and 1.24 μg/kg and 0.10-25 μg/kg for fenitrothion. The novel methods for the detection of the above mentioned organophosphorus pesticides can meet the requirements of their maximum residue limits in fruits and vegetables, which paves the way to produce rapid-test kits for the detection of organophosphorus pesticides.