食品科学 ›› 2011, Vol. 32 ›› Issue (14): 227-231.doi: 10.7506/spkx1002-6630-201114048

• 分析检测 • 上一篇    下一篇

新霉素ELISA检测方法的建立

刘沙洲1,2,桑小雪1,欧阳华学3,雷绍荣3,白林含1,*   

  1. 1. 四川大学生命科学学院
    2.成都市食品药品检测中心 3.四川省农业科学院分析测试中心
  • 出版日期:2011-07-25 发布日期:2011-06-18
  • 基金资助:
    四川省公益性研究计划项目(2008NG004)

Development of Enzyme-linked Immunosorbent Assay for Neomycin

LIU Sha-zhou1,2,SANG Xiao-xue1,OUYANG Hua-xue3,LEI Shao-rong3,BAI Lin-han1,*   

  1. (1. School of Life Sciences, Sichuan University, Chengdu 610065, China;2. Food and Drug Testing Center, Chengdu 610045, China;3. Analysis and Testing Center, Sichuan Academy of Agricultural Sciences, Chengdu 610066, China)
  • Online:2011-07-25 Published:2011-06-18

摘要: 目的:比较直接和间接竞争酶联免疫法(enzyme linked immunosorbent assay,ELISA)的优缺点,建立新霉素残留ELISA检测方法。方法:利用自制的新霉素多克隆抗体,采用直接竞争和间接竞争ELISA方法检测新霉素残留,并比较两种方法的优缺点。结果:新霉素抗血清和庆大霉素的交叉反应率为2.04%,和卡那霉素的交叉反应率为0.02%,和氨苄青霉素、红霉素、四环素的交叉反应率均小于0.01%。初步测试新霉素间接竞争ELISA法的准确性和回收率。板内误差小于4%,板间误差小于11%,回收率为135.5%~191.3%。直接竞争和间接竞争ELISA方法的检测极限分别为28.58ng/mL和51.74ng/mL,达到了国家对新霉素规定的500μg/kg MRL检测限。结论:建立了直接竞争和间接ELISA吸附检测方法,条件优化更成功的间接竞争ELISA可用于开发新霉素检测试剂盒。

关键词: 新霉素, 多克隆抗体, 竞争酶联免疫法(enzyme linked immunosorbent assay, ELISA), 方法建立

Abstract: In this study, we describe the advantages and disadvantages of direct competitive enzyme-linked immunosorbent assay (dc-ELISA) and indirect competitive ELISA (idc-ELISA) and ELISA methods for the detection of neomycin. Anti-neomycin polyclonal antibodies were prepared and used to detect neomycin by dc-ELISA and idc-ELISA. The cross-reaction rates of prepared anti-neomycin polyclonal antibodies with gentamincin and kanamycin were 2.04% and 0.02%, respectively, and with ampicillin, erythromycin and tetracycline all less than 0.01%. The accuracy and recovery of idc-ELISA were tested with an intra-plate error of less than 4%, an inter-plate error of less than 11% and a recovery between 135.5% and 191.3%. The detection limits of dc-ELISA and idc-ELISA were 28.58 ng/mL and 51.74 ng/mL, respectively, both of which were below the national maximum residue limit (MRL) of 500μg/kg. Therefore, a dc-ELISA method and an idc-ELISA method to detect neomycin have successfully established. Further, the idc-ELISA method where the working conditions were better optimized can be used for the development of neomycin test kit.

Key words: neomycin, polyclonal antibodies, competitive enzyme-linked immunosorbent assay (ELISA), method establishment

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