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| 四种牛分枝杆菌特异性蛋白融合表达及在牛结核病诊断中的临床应用 | |
| 引用本文： | 吴波,张书环,邓铨涛,项杰,陈军,刘朝,陈鑫,宋念华,陶淑珍,黄继根,韩永佩,赵波,林祥梅,陈焕春,郭爱珍.四种牛分枝杆菌特异性蛋白融合表达及在牛结核病诊断中的临床应用[J].中国奶牛,2007(10):16-20. |
| 作者姓名： | 吴波 张书环 邓铨涛 项杰 陈军 刘朝 陈鑫 宋念华 陶淑珍 黄继根 韩永佩 赵波 林祥梅 陈焕春 郭爱珍 |
| 作者单位： | 1. 华中农业大学农业微生物学国家重点实验室;华中农业大学湖北省预防兽医学重点实验室,华中农业大学动物医学院,武汉,430070 2. 湖北省武汉市结核病防治院(所),武汉,430083 3. 湖北省动物疫病诊断中心,武汉,430064 4. 武汉市动物防疫检疫站,武汉,430015 5. 武汉市奶业管理办公室,武汉,430015 6. 湖北省宜昌市夷陵区畜牧局,宜昌,443100 7. 宜昌市夷陵区奶牛养殖技术服务中心,宜昌,443100 8. 中国检验检疫科学研究院动植物检疫研究所,北京,100029 |
| 基金项目： | 国家重点基础研究发展计划(973计划);国家科技攻关项目;湖北省武汉市科技攻关项目 |
| 摘 要： | 以原核表达的MPB70-MPB83-CFP10-ESAT-6融合蛋白作为诊断抗原,建立牛结核病抗体检测间接ELISA(iELISA)。该iELISA与导致常见牛病的非相关抗原无交叉反应。用iELISA检测90份健康牛血清,确定样本阴阳性临界值(S/P)为0.17。与商品化胶体金试剂条(ICG)平行检测150份临床奶牛血清样本,结果与ICG的符合率为93.33%(140/150)。以结核菌素皮内试验(TST)为参考方法,检测了华中地区四个奶牛场的560头中国荷斯坦奶牛和90份进口奶牛结核阴性血清,结果iELISA与TST的总符合率为87.32%(489/560)。对其中22头奶牛进行鼻拭子分菌检验,4头分菌阳性奶牛的血清抗体检测也为阳性,iELISA与细菌培养的总符合率为77.27%(17/22)。以TST为参考方法,iELISA的敏感性和特异性分别为72.37%和89.67%。 |
| 关 键 词： | 融合蛋白 牛结核病 |
| 文章编号： | 1004-4264（2007）10-0016-04 |
| 收稿时间： | 2007-07-02 |
| 修稿时间： | 2007年7月2日 |
Fusion Expression of Four Proteins Specific to Mycobacterium Bovis and Its Application in Detection of Bovine Tuberculosis |
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| Wu Bo, Zhang Shuhuan, Deng Quantao, Xian Jie, Chen Jun,Liu Zhao, Chen Xin, Song Nianhua, Tao Shuzhen, Huang Jigen, Han Yongpei, Zhao Bo, Lin Xiangmei, Chen Huanchun, Guo Aizhen.Fusion Expression of Four Proteins Specific to Mycobacterium Bovis and Its Application in Detection of Bovine Tuberculosis[J].China Dairy Cattle,2007(10):16-20. | |
| Authors: | Wu Bo Zhang Shuhuan Deng Quantao Xian Jie Chen Jun Liu Zhao Chen Xin Song Nianhua Tao Shuzhen Huang Jigen Han Yongpei Zhao Bo Lin Xiangmei Chen Huanchun Guo Aizhen |
| Institution: | 1.The State Key Laboratory of Agricultural Microbiology;2.College of Veterinary Medicine,Huazhong Agricultural University,Wuhan 430070; 3.Wuhan Tuberculosis Control Institute,Wuhan 430083;4.Hubei Diagnosis Center of Animal Diseases,Wuhan 430064; 5.Wuhan Animal Disease Quarantine and Prevention Station,Wuhan 430015;6.Wuhan Dairy Administration Office,Wuhan 430015; 7.Yiling Animal Husbandry Department,Yichang 443100;8.Yiling Dairy Farming Technique Service Center,Yichang 443100; 9.Chinese Academy of Inspection and Quarantine,Institute of Animal and Plant Quarantine,Beijing 100029 |
| Abstract: | Fusion protein of MPB70-MPB83-CFP10-ESAT-6 Specific to Mycobacterium bovis was expressed and utilized as the diagnosis antigen to establish an indirect ELISA.This iELISA was demonstrated to have no cross-reaction with irrelevant antigens from other common pathogens of cows.The serum samples from 90 health cows were tested with the iELLSA and the cutoff value(S/P)was determined to be 0.17. 150 serum samples from cow herds were detected with both iELISA and colloidal gold strip test and the agreement rate reached 93.33%.Taken the tuberculin skin test as the reference test,560 serum samples from four local cow herds in the Middle of China were assayed with iELISA.As a result,the total agreement between iELISA and TST was 87.32%.22 nose swabs were collected from one of the herds to isolate Mycobacteria tuberculosis.Four cows with bacterial shedding were also positive in antibodies detected with iELISA.The agreement between bacterial isolation and iELISA was 77.2%(17/22).The sensitivity and specificity of iELISA to TST are respectively 72.3% and 89.67%.In addition,the bovine tuberculosis prevalence affected the sensitivity and specificity of iELISA. |
| Keywords: | iELISA MPB70 MPB83 CFP10 ESAT-6 |
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