猪丹毒杆菌抗体间接ELISA检测方法的建立

Development of an indirect ELISA for detection of antibodies against Erysipelothrix rhusiopathiae

为建立一种检测猪丹毒杆菌血清抗体的ELISA方法,本研究将猪丹毒杆菌(1a型)云南分离株ZY15074的超声裂解的全菌蛋白作为包被抗原,通过反应条件的优化,建立了猪丹毒杆菌抗体间接ELISA检测方法。特异性试验结果显示该方法与猪其它10种常见病原阳性血清均无交叉反应。该方法对阳性血清的敏感性为1/256。重复性试验结果显示批内、批间变异系数均小于10%。对160份临床血清样品比对试验结果显示该方法与国外间接ELISA方法的符合率为90.6%。本研究建立的间接ELISA方法具有较好的特异性、敏感性和重复性,可以用于猪丹毒杆菌血清抗体检测和流行病学调查。

To establish an indirect ELISA for detection serum antibody against Erysipelothn＇x rhusiopathiae in pigs, the indirect ELISA assay was developed using the sonicated proteins of E.rhusiopathiae strain ZY15074 （type la） isolated from Yunnan province, and the sonicated protein was used as coated protein in this study. The results of specificity test indicated that the method was specific for detecting antibodies against E.rhusiopathiae and had no cross-reactions with other pathogeny positive serum of pigs. The results of sensitivity test indicated that the detection limit for positive serum was 1/256. The results of reproducibility test showed that the coefficient of intra- and inter-variations were less than 10%. The results of comparative test showed that the coincident rate was 90.6% of the ELISA method with the imported indirect ELISA kit for detecting 160 clinical serum samples. The results indicated that the developed indirect ELISA method could be applied in serum antibodies detection and sero-epidemiology surveillance.