牛分支杆菌抗体荧光微球免疫层析试纸条检测方法的建立  

Establishment of a Fluorescent Microsphere Immunochromatographic Strip Detection Method for Mycobacterium bovis Antibody

作　　者：苏华彬 亓菲 南文龙 刘蒙达 孙明军 焉鑫 毛迎雪 曲瑶 孙淑芳 李桂梅 樊晓旭 SU Hua-bin;QI Fei;NAN Wen-long;LIU Meng-da;SUN Ming-jun;YAN Xin;MAO Ying-xue;QU Yao;SUN Shu-fang;LI Gui-mei;FAN Xiao-xu(College of Veterinary Medicine,Qingdao Agricultural University,Qingdao,Shandong 266109,China;Division of zoonoses surveillance,China Animal Health and Epidemiology Center,Qingdao,Shandong 266032,China;Key Laboratory Ruminant Infectious Disease Prevention and Control(East China),Ministry of Agriculture and Rural Affairs,Qingdao,Shandong 266032,China;College of Animal Science,Shandong Agricultural University,Taian,Shandong 271000,China)

机构地区：[1]青岛农业大学动物医学院,山东青岛266109 [2]中国动物卫生与流行病学中心,山东青岛266032 [3]农业农村部反刍动物重大疫病防控重点试验室(东部),山东青岛266032 [4]山东农业大学动物科技学院,山东泰安271000

出　　处：《动物医学进展》2024年第11期1-8,共8页Progress In Veterinary Medicine

基　　金：“十四五”国家重点研发计划项目(2022YFD1800701);国家奶牛产业技术体系资助项目(CARS36)。

摘　　要：为了建立一种便捷、高效、采用荧光微球免疫试纸条技术的牛分支杆菌检测方法,首先表达、纯化MPB70和MPB83蛋白,并用荧光微球进行标记,组装荧光微球免疫试纸条并进行条件优化、特异性试验、重复性试验以及灵敏性试验,建立一种检测牛分支杆菌的荧光微球免疫试纸条方法。试验表明,试纸条检测时间仅需15 min;试纸条仅对牛分支杆菌抗体阳性血清发生反应,特异性好;变异系数在3.51%~9.23%之间,重复性好;灵敏度与美国爱德士牛分支杆菌抗体检测试剂盒相当,且总符合率高达90.5%。说明建立的牛分支杆菌抗体荧光微球免疫层析试纸条检测方法具有良好的特异性、灵敏性与重复性,可为牛分支杆菌检测提供新技术支持。To establish a convenient,efficient Mycobacterium bovis detection method using fluorescent microsphere immunoassay strip technology,the MPB70 and MPB83 proteins were first expressed,purified,labeled with fluorescent microspheres,assembled into fluorescent microsphere immunoassay strips,and subjected to condition optimization,specificity test,repeatability test,and sensitivity test.A successful detection method for Mycobacterium bovis using fluorescent microsphere immunoassay strips was established.The experiments showed that the detection time of the strips was only 15 minutes;the strips reacted only with Mycobacterium bovis antibody-positive serum,indicating good specificity.The coefficient of variation ranged from 3.51%to 9.23%,demonstrating good repeatability.The sensitivity was comparable to the IDEXX Mycobacterium bovis antibody test kit,with an overall agreement rate of 90.5%.This indicates that the established fluorescent microsphere immunoassay strip method for detecting Mycobacterium bovis antibodies has good specificity,sensitivity,and repeatability,providing new technological support for Mycobacterium bovis diagnosis.

关 键 词：荧光微球 牛分支杆菌 试纸条 

分 类 号：S852.618[农业科学—基础兽医学] S858.23[农业科学—兽医学]