基于两个重组蛋白的牛支原体免疫胶体金抗体检测试纸条的研究  

Development of an immunocolloidal gold strip for detection of Mycoplasma bovis antibodies based on two recombinant proteins

作　　者：周华倩 储岳峰[1] 耿尚景超 郝华芳[1] 陈胜利[1] 刘永生[1] 颜新敏[1] ZHOU Hua-qian;CHU Yue-feng;GENG Shang-jing-chao;HAO Hua-fang;CHEN Sheng-li;LIU Yong-sheng;YAN Xin-min(State Key Laboratory Veterinary Etiological Biology,Lanzhou Veterinary Research Institute,Chinese Acaderrvy of Agricultural Sciences,Lanzhou 730046,China;Sichuan Agricultural University,Chengdu 611130,China)

机构地区：[1]中国农业科学院,兰州兽医研究所,家畜疫病病原生物学国家重点实验室,甘肃兰州730046 [2]四川农业大学,四川成都611130

出　　处：《中国兽医科学》2021年第8期939-946,共8页Chinese Veterinary Science

基　　金：国家重点研发计划项目(2018YFD0502305,2016YFD0500907);甘肃省重点研发计划项目(17YF1NA070);中国农业科学院兰州兽医研究所“元亨人才”计划项目(农科兰兽2020-119)。

摘　　要：本研究以牛支原体(Mycoplasma bovis)重组膜蛋白P48、P80和鸡IgY抗体标记胶体金溶液,以蛋白A和蛋白G包被T线,山羊抗鸡IgY包被C线,制备检测牛支原体抗体的胶体金免疫层析试纸条。用本实验室保存的39份牛支原体感染来源血清、56份免疫来源血清以及142份临床血清对研制的试纸条进行质量评估。结果显示,该试纸条能检测出感染和免疫14 d后的牛血清中的牛支原体抗体,对感染来源血清敏感性为95.83%、特异性为93.75%,对免疫来源血清的敏感性为92.31%、特异性为93.75%,对临床血清的敏感性为90.12%、特异性为98.36%。上述结果表明,该试纸条能用于牛支原体抗体现场快速检测、群体监测和辅助诊断。In this study,a colloidal gold immunochromatograp Hic strip was developed to detect the antibodies of Mycoplasma bovis.Colloidal gold was conjugated with the recombinant proteins P48 and P80 of M.bovis,as well as chicken’s lg Y protein,respectively.The test line was coated with proteins A and G,and the control line was coated with goat anti-chicken Ig Y.To evaluate quality of the trip,39 sera from M.bovis experimental-infected cattle,56 from M.bovis immunized cattle and 142 from field sera were employed to test.The results showed that the strip can detect antibodies against M.bovis in cattle from 14 days after challenge or immunization.The sensitivity and specificity of the strip reaches95.83% and 93.75% to experimental infection sera,92.31% and 93.75% to immune sera,90.12% and98.36% to field sera,respectively.It means that the strip can be used for rapid detection of M.bovis antibodies,which is helpful for M.bovis infection diagnosis and surveillance.

关 键 词：牛支原体 P48重组蛋白 P80重组蛋白 免疫胶体金试纸条 

分 类 号：S852.62[农业科学—基础兽医学]