检测牛结核抗体新型ELISA方法的建立及应用  

Development of a novel indirect enzyme-linked immunosorbent assay for the detection of antibodies in bovine tuberculosis

作　　者：郭设平[1] 刘思国[1] 王春来[1] 宫强[1] 邵美丽[1] 张秀华[1] 郭洋[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室/动物细菌病研究室,黑龙江哈尔滨150001

出　　处：《中国预防兽医学报》2007年第10期810-813,共4页Chinese Journal of Preventive Veterinary Medicine

摘　　要：利用分子克隆和Splicing by overlapping extension(SOE)技术,表达了重组蛋白rM70-83-E6,Western blot分析rM70-83-E6具有很好的特异性。以蛋白rM70-83-E6作为诊断抗原建立了间接ELISA方法,ELISA诊断方法的判定标准,即S/P≥0.5为阳性,S/P<0.4为阴性,0.5>S/P≥0.4为可疑。ELISA方法的特异性为96.0%、敏感性为69.4%。对67份PPD皮试阳性和50份PPD皮试阴性(117份)血清样品进行了检测,并与PPD皮试比较。67份PPD皮试阳性血清样品中有46份为ELISA检测阳性,阳性符合率为68.7%,50份PPD皮试阴性牛血清都为阴性,阴性符合率为100%,本ELISA方法与PPD皮试诊断方法总复合率为82.1%。研究表明,ELISA方法具有很好的开发和应用前景。Enzyme-linked immunosorbent assay （ELISA） for diagnosis of bovine tuberculosis has been widely explored. The fusion protein M70-83-E6 （rM70-83-E6） expressed in E.coli was shown specifically react to sera from cattle with bovine tuberculosis. In this work, an indirect ELISA was established using M70-83-E6 protein as the diagnostic antigen. The ELISA were nomalised with 150 serum samples from healthy cattle, with a S/P ≥ 0.5 being positive and S/P 〈0.4 negative, Test of 85 serum samples from cows with bovine tuberculosis and 100 serum samples from healthy cattle detected 69.4 % positive with a specificity of 96,0 %. The ELISA results of 117 serum samples （67 from bovine with positive skin test of PPD; 50 from bovine with negative skin test of PPD） were compared with those of skin test of PPD. 46 out of the 67 samples from bovine with positive skin test of PPD were also positive by ELISA, with a positive coincidence of 68.7 %. All 50 samples from bovine with negative bovine skin test of PPD were also negative by ELISA. The total coincidence of the ELISA method and skin test of PPD was 82.1%.

关 键 词：融合蛋白 牛结核病 间接ELISA 

分 类 号：S854.43[农业科学—临床兽医学]