食品科学 ›› 2009, Vol. 30 ›› Issue (20 ): 416-420.doi: 10.7506/spkx1002-6300-200920094

• 分析检测 • 上一篇    下一篇

氯霉素免疫检测方法的建立

胡拥明,梁沙莉,徐丽广,陈 伟,王利兵,胥传来*   

  1. 江南大学食品学院
  • 收稿日期:2009-07-08 出版日期:2009-10-15 发布日期:2010-12-29
  • 通讯作者: 胥传来

Development of Immunoassay for Detection of Chloramphenicol

HU Yong-ming,LIANG Sha-li,XU Li-guang,CHEN Wei,WANG Li-bing,XU Chuan-lai*   

  1. School of Food Science and Technology, Jiangnan University,Wuxi 214122, Chian
  • Received:2009-07-08 Online:2009-10-15 Published:2010-12-29
  • Contact: XU Chuan-lai

摘要:

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%。

关键词: 氯霉素, 间接竞争酶联免疫, 残留

Abstract:

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.

Key words: chloramphenicol, IC-ELISA, residue

中图分类号: