牛副结核胶体金免疫层析试纸条的研制  

Development of collcidal gold immunochromatographic test strip for detection of bovine paratuberculosis

作　　者：乔榛[1] 张丽娟[1] 秦建华[1] 赵月兰[2] 左玉柱[1] 

机构地区：[1]河北农业大学动物科技学院,河北保定071001 [2]河北农业大学中兽医学院,河北定洲073000

出　　处：《畜牧与兽医》2010年第1期5-7,共3页Animal Husbandry & Veterinary Medicine

基　　金：国家"十一五"科技支撑计划(2006BAD04A10-4);河北省重大技术创新专项计划(07227146Z-4)

摘　　要：将提纯的牛副结核分支杆菌重组蛋白MAP0862-2154c作为硝酸纤维素膜检测线的包被抗原,兔抗羊IgG作为硝酸纤维素膜质控线的包被抗体,金标羊抗牛IgG点喷到玻璃纤维素膜上,制成用于检测牛副结核抗体胶体金免疫层析试纸条。用牛的副结核标准阳性血清与检测试纸条反应,在检测线处出现红色反应带,而滴加阴性血清的试纸条检测线处未出现反应条带,上述2种血清在质控线处均出现红色反应条带;试纸条可检出牛副结核抗体的最低抗原包被浓度为400μg/mL;试纸条不与牛结核病、牛布鲁氏菌病的阳性血清发生反应;试纸条37℃保存9d后的检验结果与4℃保存的检验结果相同。所制备的试纸条具有灵敏、特异的优点,稳定性良好;10min左右即出结果,操作简便,可用于临床诊断。The purified recombinant protein of Mycobacterium paratuberculosis, MAP0862-2154c, was used as coated antigen at the detected line position of nitrocellulose membrane, while rabbit anti-sheep IgG was used as antibody at the control line position of nitrocellulose membrane, and colloidal gold labeled goat anti-bovine IgG was sprayed on glass cellulose membrane, thus to prepare the colloidal gold immunochromatograpic strip. A red reaction strip at the test line position appeared only when the standard positive serum was used. The red reac- tion strips appeared at the control line position whatever sera were used. The minimum concentration of coated antigen was 400 ug/mL. The strips did not react with positive sera of bovine tuberculosis and bovine brucellosis. The detection results of different batches of test strips stored in 37℃ for 9 d were the same as those in 4℃. The results revealed that the test strips has good sensitivity, specificity and stability. The technique is easy to operate and can be used-for clinical diagnosis.

关 键 词：牛副结核 胶体金 免疫层析 试纸条 

分 类 号：S858.23[农业科学—临床兽医学]