氯霉素免疫检测方法的建立

采用碳二亚胺法(EDC)和混合酸酐法，分别与牛血清白蛋白(BSA)和卵清白蛋白(OVA)偶联制备氯霉素免疫原CAP-HS-BSA]和包被原(CAP-HS-OVA)。经紫外和变性电泳(SDS-PAGE)鉴定，免疫原与包被原均制备成功，偶联比分别为1:16和1:17。利用免疫原免疫两只新西兰大耳白兔，制备多克隆抗体，建立间接竞争ELISA检测氯霉素残留的方法。建立的ic-ELISA检测氯霉素方法的半数抑制浓度(IC50)为7.275ng/ml，通过对氯霉素琥珀酸钠和链霉素、青霉素及庆大霉素的交叉反应率的测定，结果显示抗体对氯霉素琥珀酸钠有较高的交叉反应率而对链霉素、青霉素和庆大霉素的交叉反应率均小于0.1%。

Development of Immunoassay for Detection of Chloramphenicol

Immunogen and coating antigens of chloramphenicol were successfully synthesized through coupling with BSA and OVA using the method of EDC and mixed anhydride. The conjugation was evaluated using UV spectroscopy and SDS -PAGE, and the conjugation rates were estimated to be 1:17 and 1:16, respectively. Two New Zealand rabbits were immuned by CAP-HS-BSA to generate polyclonal antibody. The titers of both antibodies were 218700 and 72900, respectively, which reached the standard of enzyme linked immunosorbent assay (ELISA). An indirect competitive ELISA was developed to determine chloramphenicol using these antibodies. The IC50 for chloramphenicol was 0.86 μg/L using ELISA determination. The specificity of chloramphenicol sodium succinate, streptomycin, penicillin and gentamicin was evaluated using cross reactivity, which revealed that the developed antibody had high specificity to chloramphenicol sodium succinate, while low specificity to other three drugs.