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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。
A chemiluminescence enzyme immunoassay (ELISA) was developed for the simultaneous determination of three organophosphorus pesticide residues using a broad spectrum of specific monoclonal antibodies that recognize parathion, methyl parathion and fenitrothion simultaneously. , CLEIA). Two kinds of reaction modes, ic-CLEIA and dc-CLEIA, were compared. The optimal physicochemical parameters were optimized and the optimal reaction conditions were established. The results showed that the medium inhibitory concentration (IC 50) of indirect competition CLEIA assay for parathion, methyl parathion and fenitrothion were 5.57, 2.30 and 2.62 μg / kg, respectively. The linear range of detection was 0.39 ~ 100, 0.39 ~ 25 and 0.39 ~ 25μg / kg, respectively. The direct inhibition of CLEIA method for the determination of parathion, methyl parathion and fenitrothion in the concentration of inhibition (IC 50) were 5.43,1.34 and 1.24μg / kg. The linear range was 0.39-100, 0.10-25 and 0.10-25 μg / kg, respectively. The established CLEIA method can basically meet the testing requirements of the maximum residue limits of parathion, methyl parathion and fenitrothion in cereals and fruits and vegetables, providing the technical basis for the development of multi-residue detection kit of organophosphorus pesticides.