Establishment of double sandwich ELISA for detecting infectious bursal disease virus

Immune sera of rabbit/mouse anti-infectious bursal disease virus(IBDV) and sheep anti-rabbit were prepared,and immunoglobulins(IgGs) were purified from the three immune sera and labeled with HRP.Then,the optimal working concentrations of the IgGs were determined by using the chessboard titration crossing test and double sandwich ELISA based on the IgGs was established for the detection of IBDV.The results showed that the optimal working concentration of the IgGs was 1∶160,the optimal dilution of rabbit anti-IBDV IgG was 1∶200 and the optimal dilution of sheep anti-rabbit antibody labeled by HRP was 1∶400.Double sandwich ELISA had high specificity and reproducibility,which was confirmed by the repeating,blocking and crossing tests and it had no cross-reaction with Newcastle disease virus(NDV),infectious bronchitis virus(IBV) and eggs drop syndrome virus(EDSV).It was confirmed by the diagnosis of the experimental cases that the established method could be used for rapid diagnosis of clinical IBDV.