新霉素ELISA检测方法的建立  

Development of Enzyme-linked Immunosorbent Assay for Neomycin

作　　者：刘沙洲[1,2] 桑小雪[1] 欧阳华学[3] 雷绍荣[3] 白林含[1] 

机构地区：[1]四川大学生命科学学院,四川成都610065 [2]成都市食品药品检测中心,四川成都610045 [3]四川省农业科学院分析测试中心,四川成都610066

出　　处：《食品科学》2011年第14期227-231,共5页Food Science

基　　金：四川省公益性研究计划项目(2008NG004)

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。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. Antineomycin 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 500 pg/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.

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 

分 类 号：S854[农业科学—临床兽医学]