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目的:制备抗有机氯农药DDT代谢物DDA的多克隆抗体,从而建立DDA的ELISA检测方法,并将其初步应用于贝类样品中的DDA检测。方法:采用碳二亚胺法制备DDA-BSA完全抗原,对兔进行免疫,得到兔抗DDA多克隆抗体,并应用于建立DDA的间接酶联免疫检测方法(Enzyme Linked Immunosorbent Assay,ELISA)。结果:成功制备抗DDA多克隆抗体,其IC50为115.30 ng/ml,最低检出限(IC20)为20.88 ng/ml,回收率为90.05%～106.38%,批内变异系数为1.25%～6.77%。结论:制备的抗DDA多克隆抗体及建立的ELISA检测方法可用于贝类样品中DDA含量的检测。
OBJECTIVE: To prepare polyclonal antibodies against DDT metabolites of organochlorine pesticides (DDT), and to establish an ELISA method for the determination of DDA. The method was initially applied to the detection of DDA in shellfish samples. Methods: The complete antigen of DDA-BSA was prepared by carbodiimide method, and the rabbit anti-DDA antibody was obtained. The polyclonal antibody against rabbit polyclonal antibody was obtained and used to establish the indirect enzyme-linked immunosorbent assay (ELISA) of DDA. Results: The anti-DDA polyclonal antibody was successfully prepared with the IC50 of 115.30 ng / ml, the IC 20 of 20.88 ng / ml and the recovery of 90.05% -106.38%. The intra-assay CV was 1.25% -6.77% . Conclusion: The prepared anti-DDA polyclonal antibody and established ELISA detection method can be used for the determination of DDA content in shellfish samples.