牛支原体抗体胶体金免疫层析检测方法的建立及应用  

Development and application of a colloidal gold immunochromatographic assay for detection of antibodies against Mycoplasma bovis

作　　者：田可昕 高鹏程[2] 陈胜利[2] 郑福英[2] 许健 付强[1] 史慧君[1] 储岳峰[2] TIAN Ke-xin;GAO Peng-cheng;CHEN Sheng-li;ZHENG Fu-ying;XU Jian;FU Qiang;SHI Hui-jun;CHU Yue-feng(College of Veterinary Medicine,Xinjiang Agricultural University,Urumqi 830052,China;State Key Laboratory for Animal Disease Control and Prevention/College of Veterinary Medicine of Lanzhou University/Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou 730046,China)

机构地区：[1]新疆农业大学,动物医学学院,新疆乌鲁木齐830052 [2]中国农业科学院兰州兽医研究所兰州大学动物医学与生物安全学院,动物疫病防控全国重点实验室,甘肃兰州730046

出　　处：《中国兽医科学》2023年第7期831-836,共6页Chinese Veterinary Science

基　　金：国家重点研发计划项目(2022YFD1302101);中国农业科学院科技创新工程(CAAS-ASTIP-2021-LVRI);甘肃省重点人才项目(2021RCXM04)。

摘　　要：本试验以牛支原体(Mycoplasma bovis)MilA基因重组蛋白P300为检测靶标抗原包被检测线(T线),山羊抗家兔IgG包被质控线(C线),金黄色葡萄球菌蛋白A(SPA)标记胶体金溶液,以建立牛支原体抗体胶体金免疫层析试纸条检测方法。通过检测牛布氏杆菌、丝状支原体、牛病毒性腹泻病毒、牛巴氏杆菌和口蹄疫病毒5种病原的阳性血清以及实验室保存的牛支原体阳性血清和临床血清,对该方法的特异性、敏感性和重复性进行了验证。结果显示,该方法检测牛布氏杆菌、丝状支原体、牛病毒性腹泻病毒、牛巴氏杆菌和口蹄疫病毒5种病原的阳性血清均为阴性;对已知牛支原体阳性血清的检测最高稀释梯度可达1∶2 048;对临床血清检测,与ELISA的检测结果符合率为94.04%。结果表明,牛支原体抗体胶体金免疫层析试纸条特异性好、敏感性高。可用于牛支原体感染的快速诊断,尤其适用于现场检测。A rapid detection method for Mycoplasma bovis antibodies using a colloidal gold im-munochromatographic test strip has been established.The detection target antigen is the M.bovis MilA gene recombinant protein P300,which is coated on the test line(T line).The quality control line(C line)is coated with goat anti-rabbit IgG,and the colloidal gold solution is labeled with Staphylococcus au-reus protein A(SPA).To verify the specificity,sensitivity,and repeatability of this method,positive sera for Brucella bovis,M.mycoides,bovine viral diarrhea virus,Pasteurella bovis,orfoot-and-mouth disease virus were detected,as well as positive sera of M.bovis preserved in the laboratory and clinical sera.In result,this method accurately detected M.bovis antibodies,while the positive sera forthe other pathogens were all negative.The highest dilution gradient of positive M.bovis serum was 1∶2048,and the coincidence rate with ELISA detection of clinical serum was 94.04%.The results showed that the colloidal gold immunochromatographic test strip for detecting M.bovis antibodies has good specificity and high sensitivity,and is suitable for rapid diagnosis of M.bovis infections,especially for on-site detection.

关 键 词：牛支原体 P300重组蛋白 胶体金免疫层析 

分 类 号：S852.62[农业科学—基础兽医学]