牛布鲁氏菌抗体荧光快速检测试纸卡的研制  

Development of a Rapid Diagnostic Kit for Bovine Brucella Antibodies

作　　者：王英超 赵荣茂 张慧莹 刘海莹 高晓龙 高敏 郑博君 周德刚 梅力 冯小宇 Wang Yingchao;Zhao Rongmao;Zhang Huiying;Liu Haiying;Gao Xiaolong;Gao Min;Zheng Bojun;Zhou Degang;Mei Li;Feng Xiaoyu(Beijing Center for Animal Disease Control and Prevention,Beijing 102629,China;NBGen Biotechnology Co.,Ltd.,Beijing 100176,China)

机构地区：[1]北京市动物疫病预防控制中心,北京102629 [2]北京纳百生物科技有限公司,北京100176

出　　处：《中国动物检疫》2023年第2期125-130,152,共7页China Animal Health Inspection

基　　金：2021年北京市农业农村局农业科技项目(20210112)。

摘　　要：为研制一种快速检测牛布鲁氏菌抗体的便捷试纸卡,采用间接荧光免疫层析技术,以荧光微球为标记物,与兔抗牛IgG偶联,以布鲁氏菌脂多糖(LPS)抗原包被硝酸纤维素膜,通过反应条件优化,研制牛布鲁氏菌抗体荧光微球快速检测试纸卡。结果显示:该试纸卡灵敏度可达0.8 IU/mL,与牛口蹄疫病毒O型、牛口蹄疫病毒A型、牛病毒性腹泻病毒、牛传染性鼻气管炎病毒的抗体阳性血清均无交叉反应;对469份临床牛血清进行检测,同时与荧光偏振方法进行比较,发现两种方法的符合率为100%。结果表明,本次研制的牛布鲁氏菌抗体荧光微球检测试纸卡灵敏度高,特异性好,适用于临床样品的快速检测。本试纸卡的成功研制为牛布鲁氏菌病免疫效果评估和准确诊断提供了一种简便的血清学诊断方法。In order to develop a rapid test strip for bovine Brucella antibodies,a fluorescent microsphere was used as a marker and coupled with rabbit anti-bovine IgG by the indirect fluorescence immunochromatography,then the nitrocellulose filter membrane was enveloped by Brucella lipopolysaccharide(LPS)antigen,followed by optimization of reaction conditions,and such a kit was developed.The results showed that,the sensitivity of the kit was up to 0.8 IU/mL,and failed to react with the antibody positive sera of bovine foot-and-mouth disease virus type O(FMD-O),bovine foot-and-mouth disease virus type A(FMD-A),bovine viral diarrhea virus(BVDV)and bovine infectious rhinotracheitis virus(IBRV);469 clinical bovine serum samples were tested by the kit that was then compared with the fluorescence polarization method,it was found that the coincidence rate of the two methods was 100%.In conclusion,the kit developed in the study was with high sensitivity and specificity,which was suitable for rapid detection for clinical samples.A rapid serological diagnostic method was therefore provided for evaluation on the immune effect and accurate diagnosis of bovine brucellosis.

关 键 词：牛布鲁氏菌 荧光免疫层析技术 试纸卡 荧光微球 脂多糖抗原 

分 类 号：S855.12[农业科学—临床兽医学]