诊断抗原的纯化在牛γ-干扰素ELISA法中的应用  

Purification in antigen and application in diagnosis of IFN-γ-ELISA detection of bovine tuberculosis

作　　者：邱美珍[1] 周望平[1] 肖兵南[1] 杜丽飞[1] 胡述光[2] 刘毅[2] 陈江[2] 

机构地区：[1]湖南省畜牧兽医研究所,湖南长沙410131 [2]湖南农业大学动物医学院,湖南长沙410128

出　　处：《湖南农业大学学报（自然科学版）》2011年第1期82-85,共4页Journal of Hunan Agricultural University(Natural Sciences)

基　　金：国家科技支撑计划项目(2006BAD04A15-2)

摘　　要：比较皮内变态反应(TST)、抗酸染色、牛结核ELISA(PPD-ELISA)和γ-干扰素ELISA法(IFN-γ-ELISA)对牛结核的检测结果,并通过比较牛PPD、禽PPD、亲和层析牛PPD(牛PPD’)的IFN-γ-ELISA检测结果,探索诊断抗原的纯化在牛IFN-γ-ELISA法中的应用.结果表明,与抗酸染色相比,IFN-γ-ELISA的敏感性、特异性和符合率都比较高,分别为83.33%(5/6),98.14%(53/54),98.33%(58/60);牛PPD、禽PPD、牛PPD’3种特异性抗原刺激产生IFN-γ释放反应的结果表明,1例抗酸染色阳性的牛,未经亲和层析处理牛PPD的IFN-γ-ELISA检测OD值偏高,判为牛结核阳性,亲和层析纯化牛PPD的IFN-γ-ELISA检测OD值较低,判为牛结核阴性,表明亲和层析处理能排除环境分支杆菌对试验的干扰,IFN-γ-ELISA法有望替代TST法在中国推广使用。The test results of the bovine tuberculin skin test（TST）,acid-fast stain test,ELISA test and bovine IFN-γ-ELISA diagnosis test respectively were compared for detection of tuberculosis of them,three stimulators,detected by IFN-γ-ELISA including bovine PPD,avian PPD and bovine PPD at affinity chromatography were compared in exploration for application of purification of diagnostic antigen in the IFN-γ-ELISA method.Using acid-fast staining as the standard,the sensitivity of IFN-γ-ELISA detection reached 83.33%（5/6）,and its specificity reached 98.14%（53/54）,the consistency reached 98.33%（58/60）.As the other result,acid-fast stain detected a positive sera with high OD value and produced a positive result using bovine IFN-γ-ELISA in which the antigen was not purified at affinity chromatography,but the result was negative with low OD value using bovine IFN-γ-ELIS in which the antigen was purified at affinity chromatography.In conclusion,the miscarriage of justice caused by infected with Mycobacterium bovis and environmental Mycobacterium simultaneouly could be prevented using bovine IFN-γ-ELISA in which the antigen was purified at affinity chromatography.So this method has great potential to replace TST for development and application prospects in China.

关 键 词：牛结核 皮内变态反应 抗酸染色 牛结核ELISA γ-干扰素ELISA法 亲和纯化 

分 类 号：S858.23[农业科学—临床兽医学]