牛结核胶体金免疫层析快速诊断方法的建立及初步应用

Establishment and Preliminary Application of Immunochromatographic Strips for Rapid Detection of Antibodies Against M.bovis

建立一种快速检测牛结核抗体的方法,用以诊断牛结核病。根据胶体金免疫层析技术原理,用兔抗牛血清蛋白和大肠埃希菌表达的牛分支杆菌MPB70-83-ESAT-6蛋白分别作为胶体金标记抗原和检测线上的捕获杭原,制备牛结核抗体检测试纸条。结果表明,MPB70-83-ESAT-6抗原的最适包被浓度为2.5mg/mL;研制的试纸条不与牛的其他疾病的阳性血清反应,具有较高的特异性;该试纸条的最高检出稀释度为1∶640;试纸条与TST的符合率为80%。该试纸条具有敏感、特异、简便、快速的特点,适用于对牛结核病进行普查和检疫,具有良好的应用前景。

To develop a new method to rapidly detect antibodies against Mycobacterium boris for diagnosis of bovine tuberculosis（bTB）. Strips were prepared with the principle of colloidal gold immunochromato graphy assay（GICA）. Rabbit anti-bovine serum was labeled by colloidal gold and MPB70-83-ESAT-6 was used as the test capture reagent to detect the corresponding antibodies. The optimal concentration of capture reagent MPB70-83 -ESAT 6 was 2.5 mg/mL. The strips did not react with positive sera of other bovine diseases and displayed high specificity. The maximum detection dilution of the strips was 1 ： 640 in detecting clinical samples, Comparied with tuberculin skin test （TST）,the agreement ratios were 80%. This novel colloidal gold test strip for bTB antibody detection is sensitive, specific, convenient to be used, rapid to get results. It will be a good method in bTB survey and in bTB eradication program.