Abstract: An indirect ELISA was established to detect antibody against duck hemorrhagic disease virus (DHDV) using purified DHDV as coating antigen.The reaction conditions were optimized, including 7.06 μg/well coating antigen of the virus,1:25 dilution of testing serum and 1:500 dilution of HRP conjugated anti-duck IgG with cutoff-value of 0.44 (D_{450 nm}).The specific tests showed that there were no cross-reaction to the anti-serum against duck plague virus, duck hepatitis virus, Muscovy duck parvovirus, Muscovy duck reovirus, which showed that the ELISA was specific to anti-serum against DHDV. The intro-and inter-assay indicated that the coefficient of maximum variation was 0.0221 and 0.0032, respectively, which showed that the method had good stability. And the result was the same as the neutralization test. The method was proved to be rapid,specific and repetitive by detecting different serum samples from immune ducks, which might be used as a serodiagnosis of DHDV.

Key words: DHDV; indirect ELISA; antibody