猪化脓隐秘杆菌抗体间接ELISA检测方法的建立

Establishment of an indirect ELISA for detection of swine antibodies against Trueperella pyogenes

为建立检测化脓隐秘杆菌(T.pyogenes)血清抗体的间接ELISA方法,本研究以超声破碎处理T.pyo-genes分离株TP-2849(NZ_CP029004)的全菌蛋白作为包被抗原,经反应条件优化建立了T.pyogenes抗体间接ELISA检测方法。该方法与7种常见病原阳性血清均无交叉反应,特异性较强;敏感性试验显示阳性血清的检测下限为1∶128,敏感性较高;组内、组间变异系数均小于9.5%,重复性较好。对吉林省某养殖场100份猪血清检测结果显示,该方法与PCR检测方法总符合率为94%。本研究建立的间接ELISA方法为T.pyogenes抗体检测提供了一种新手段。

To establish an indirect ELISA(iELISA)for detecting antibodies against Trueperella pyogenes,we used the sonicated lysate of T.pyogenes TP-2849 strain(NZ_CP029004)as the coating antigen and optimized reaction conditions.The specificity test result showed that the iELISA had no cross-reactions with the other seven common pathogenic positive sera of H.parasuis,E.coli,P.multocida,CSFV,PRRSV,PPV and PRV,which showed the established assay was high specificity for detection of the antibody against T.pyogenes.The detection limit of the iELISA for positive serum was 1∶128.The coefficient of intra-variations and inter-variations were both less than 9.5%.Above all,the established iELISA provide a new method for detection of the antibody against T.pyogenes.