摘要：

Chemical modification method was employed to synthesize the thiamethoxam hapten,which was then coupled with BSA and ovalbumin(OVA) to prepare immunogen and coating antigen through EDC method,respectively.And satisfied molecule coupling ratios between the thiamethoxam hapten and carrier proteins were eventually obtained.In order to obtain the monoclonal antibodies against thiamethoxam,the 6-week old Balb/c mice were immunized with the prepared immune antigen,and cells hybridization,positive screening and cloning.The related experimental results showed that the titer of the monoclonal antibodies was reached 1∶6.4×105 and the antibody subclass was all IgG1.After optimization of the ELISA conditions,an indirect competitive ELISA for quantitative analysis of thiamethoxam was established.The limit of detection(IC10),limit of quantification(IC20) and median inhibitory concentration(IC50) were 0.001 mg/L,0.0022 mg/L and 0.0255 mg/L,respectively.The produced monoclonal antibodies had no cross-reaction with some other analogues of thiamethoxam except for Lothianidin.The recovery rates were determined in Xinxiang city tap water spiked at 0.01 mg/L,0.5 mg/L and 10 mg/L.The overall recovery rates were over 75%,with a relative standard deviation(RSD) less than 8%.This developed method has the characteristics of high accuracy and precision,and can used to determine the thiamethoxam residue in water.

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