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.
作者
张泽宇
苏扣
王瑞
霍乃蕊
赵思俊
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)
出处
《中国动物检疫》
CAS
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)
作者简介
通信作者:霍乃蕊;通信作者:赵思俊.
