三甲氧苄氨嘧啶单克隆抗体制备以及酶联免疫试剂盒的研究

Study on the Production of Trimethoprem Monoclonal Antibodies and Its ELISA Kit for Rapid Detection

[目的]探讨系统地检测水质中三甲氧苄氨嘧啶残留量的方法。[方法]通过三甲氧苄氨嘧啶与马来酸酐反应，得到三甲氧苄氨嘧啶半抗原，再通过免疫动物得到抗三甲氧苄氨嘧啶单克隆抗体，并将其应用于能够检测水质中三甲氧苄氨嘧啶残留量的ELISA试剂盒。[结果]试验表明，该试剂盒对水质中三甲氧苄氨嘧啶的检测限为2．34μg／kg，IC50（50％抑制浓度）为4．8μg／L，回收率为60．5％～79．7％，试剂盒的标准曲线范围为0～80μg／L，批内、批间的相对标准偏差均小于10％，三甲氧苄氨嘧啶单克隆抗体与二甲氧苄氨嘧啶的交叉反应率小于1％，4℃下能够保存12个月，稳定性较好。[结论]研究可为监管三甲氧苄氨嘧啶的滥用提供参考。

Objective] The aim was to discuss the method for detecting Trimethoprem residues in water.[Method] The synthesized Trime-thoprem hapten was prepared from a sequence of reactions that used Trimethoprem and Maleic anhydride .And Trimethoprem monoclonal anti-bodies were prepared by animal immune.And ELISA kit was prepared.Then the limit of detection and relative standard deviation（RSD） of trimethoprem ELISA was studied.[Result] The limit of detection was 2.34 μg/kg, with the IC50 values 4.8 μg/L.The recoveries were be-tween 60.5%-79.7%.The standard curve ranged from 0 to 80 μg/L, and RSD was less than 10%.The cross reaction rate with Diaveridine was less than 1%.The stability tests results revealed that the kit can be kept at 4 ℃ for 12 months.[ Conclusion] The study can provide ref-erence for administration of the abuse of Trimethoprem.