ELISA测定猪尿液中β-受体激动剂残留的准确性评价  

Determination of β-receptor Agonist Residue in Pig Urine by ELISA

作　　者：张泽宇 苏扣 王瑞 霍乃蕊[1] 赵思俊[2] Zhang Zeyu;Su Kou;Wang Rui;Huo Nairui;Zhao Sijun(College of Animal Science and Veterinary Medicine,Shanxi Agricultural University,Taigu,Shanxi 030801,China;China Animal Health and Epidemiology Center,Qingdao,Shandong 266032,China;College ofAnimal Science and Veterinary Medicine,Tianjin Agricultural University,Tianjin 300384,Chinap;College ofAnimal Science and Veterinary Medicine,Anhui Agricultural University,Hefei,Anhui 230036,China)

机构地区：[1]山西农业大学动物科技学院,山西太谷030801 [2]中国动物卫生与流行病学中心,山东青岛266032 [3]天津农学院动物科学与动物医学学院,天津300384 [4]安徽农业大学动物科技学院,安徽合肥230036

出　　处：《中国动物检疫》2019年第5期103-109,共7页China Animal Health Inspection

基　　金：国家畜禽产品风险评估专项(GJFP201800703)

摘　　要：本研究建立克伦特罗间接竞争ELISA检测方法,对检测条件进行了优化确定,并利用猪尿液进行添加回收试验。结果显示:ELISA反应CLE-OVA包被抗原和CLE单抗的最适浓度分别为1:40 000和1:20 000,间接竞争ELISA的线性范围为0.016 l^8.520 0 ng/mL。该方法对克伦特罗、溴氯特罗、马布特罗和溴布特罗均有交叉反应。对冷冻储存半年以上的猪尿进行ELISA测定时,发现检测限可达58.848 3 ng/mL,药物添加回收率为85.19%~5 173.25%,变异系数为4.78%~96.79%;而经过MCX固相萃取柱净化处理后,检测限为0.076 ng/mL,添加回收率在101.21%~119.10%之间,变异系数为6.05%~18.38%。结果表明,SPE净化处理后,可有效去除尿液中的色素、蛋白等基质,提高了该法的灵敏度和准确性。In this study,an indirect competitive enzyme-linked immuno-sorbent assay(ic-ELISA)for detection of Clenbuterol was established through optimizing all detection conditions and recovery test by use of porcine urine. The results showed that the optimal concentration of coating antigen(CLE-OVA)and monoclonal antibody (CLE)of ELISA reaction was 1:40 000 and 1:20 000 respectively,the linear range of indirect competitive ELISA was 0.016 l–8.520 0 ng/mL. By the method,the cross-reaction occurred with Clenbuterol,Bromoclonbuterol, marbuterol and brombuterol. When porcine urine stored for more than half a year was determined by ELISA,it was found that the detection limit reached 58.848 3 ng/mL,the drugs added recovery was 85.19% to 5 173.25% and the variable coefficient was 4.78% to 96.79%.While the detection limit was 0.076 ng/mL,the added recovery ranged within 101.21% to 119.10% and the variable coefficient remained 6.05% to 18.38% after purified by MCX solid phase extraction column(MCX-SPE). It was concluded that the matrix,such as pigments,proteins,etc.,could be effectively removed from porcine urine after purified by SPE,which improved the sensitivity and accuracy of the method.

关 键 词：克伦特罗 猪尿 间接竞争ELISA 固相萃取 

分 类 号：S851.347[农业科学—预防兽医学]