牛布鲁氏菌BP26间接ELISA检测方法的建立

Development of indirect ELISA for Brucella abortus coating with BP26

为建立检测牛布鲁氏菌(Brucella)血清抗体的间接ELISA方法,本研究将原核表达并纯化的Brucella外膜蛋白BP26作为包被抗原,采用方阵法确定包被抗原和待检血清的最佳工作浓度,经过对各种反应条件进行优化,建立了牛Brucella间接ELISA抗体检测方法。该方法对其他相关的牛病病原无交叉反应,其组内和组间的变异系数均小于10%,具有良好的重复性。对100份临床血清样品进行检测,同时将建立的间接ELISA方法与IDEXX公司的同类试剂盒进行比较,符合率为93%。本研究为布鲁氏菌病提供了一种简便的血清学诊断方法。

To develop rapid method for detecting antibodies against Brucella abortus, an indirect enzyme-linked immunosorbent assay （iELISA） was established using the purified recombinant protein BP26 expressed in E.coli as coating antigen. Under the optinal conditions including working concentrations of coating antigen and serum samples. The results showed that the assay was specific to detect the antibody against B.abortus in bovine serum and no cross-reaction with positive antiserum from other related bovine diseases. Furthermore, comparing with the commercial ELISA kit form IDEXX company, the specificity was 92%, sensitivity was 94% and coincidence rate was 93%. The established iELISA method provided a simple and rapid serological diagnostic.