鹿布鲁菌病免疫胶体金检测试纸条的研制  

Development of a colloidal gold immunochromatographic test strip for detecting Brucellosis in deers

作　　者：郭健[1] 张月 王 舸 孙嘉慧 李艳[1] 杨艳玲[2] GUO Jian;ZHANG Yue;WANG Yange;SUN Jiahui;LI Yan;YANG Yanling(Jilin Medical University,Jilin 132013,China;State Key Laboratory for Molecular Biolgy of Special Economic Animals/Institute of Special Animal and Plant Sciences,Chinese Academy of Agricultural Sciences,Changchun 30122,China)

机构地区：[1]吉林医药学院,吉林吉林132013 [2]中国农业科学院特产研究所特种经济动物分子生物学重点实验室,吉林长春130122

出　　处：《畜牧与兽医》2019年第11期132-136,共5页Animal Husbandry & Veterinary Medicine

基　　金：吉林省抗体工程科技协同创新中心资助项目(20180623045TC);十三五重点研发专项(2016YFD00500900);吉林省卫生厅科研计划项目(2015Z041)

摘　　要：利用SUMO-BCSP31重组大肠杆菌表达并纯化了布鲁菌BCSP31重组蛋白,并利用该蛋白研制了布鲁菌免疫胶体金检测试纸条。以BCSP31蛋白作为检测抗原,通过间接法研制胶体金试纸条检测鹿血清抗体。该试纸条与大肠杆菌、沙门菌、小肠结肠炎耶尔森菌和绿脓杆菌无交叉反应;检测阳性血清最大稀释倍数为1∶640,证明试纸条具有良好的特异性和敏感性。临床上对420只鹿进行检测,比对试纸条与虎红平板凝集试验(RBT)检测结果:阳性符合率为100%,阴性符合率为96%,Kappa值为0.73,两种方法符合率较高;比对试纸条与cELISA检测结果:阳性符合率为81%,阴性符合率为100%,Kappa值为0.88,两种方法符合率高。该试纸条简便快捷,具有较高准确性,可用于鹿布鲁菌病的现场初筛检测。In this study, the recombinant protein of SUMO-BCSP31 was expressed and purified in an E. coli expression system, and then a colloidal gold-labelled immunochromatographic test strip of Brucellosis targeting at BCSP31 protein was developed to detect antibodies in deer serum. The results showed that the test strips did not cross-react with the antibodies against E. coli, Salmonella, Yersinia enterocolitica and Pseudomonas aeruginosa;and the maximum dilution of detectable serum was 1∶640. This result indicated that the test strips had good specificity and sensitivity. A total of 420 clinical deer were tested in this study through the strip and RBT tests which showed that the positive coincidence rate was 100%, the negative coincidence rate was 96%, and the Kappa value was 0.73. The two methods both had a high coincidence rate. The results of the strip and cELISA tests showed that the positive coincidence rate was 81%, the negative coincidence rate was 100%, and the Kappa value was 0.88. The coincidence rate of the two methods was higher. The test strip developed in this study had high accuracy and might be used for convenient quick on-site screening detection of brucellosis in deer.

关 键 词：布鲁菌 BCSP31 免疫层析 胶体金试纸条 

分 类 号：S858.25[农业科学—临床兽医学]