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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
【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。
【Abstract】 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.
【Key words】 bovine tuberculosis; tuberculin skin test(TST); acid-fast stain; PPD-ELISA; IFN-γ ELISA; affinity chromatography;
- 【文献出处】 湖南农业大学学报(自然科学版) ,Journal of Hunan Agricultural University(Natural Sciences) , 编辑部邮箱 ,2011年01期
- 【分类号】S858.23
- 【被引频次】1
- 【下载频次】87