Abstract: In order to explore the feasibility of DF-1 cells replacing chicken embryo fibroblast（CEF） cells as a culture substrate for the determination of the antigen contents of Infectious bursal disease virus（IBDV） liquid and serum neutralizing antibody titer, in the experiment, DF-1 cells and CEF cells were used as culture substrates to determine the antigen contents of six batches of IBDV BJQ902 strain virus liquid, and neutralizing antibody titers of SPF chickens immunized with different doses of ND-IB-IBD（La Sota strain+M41 strain +BJQ902 strain） triple inactivated vaccine and serum of non immunized SPF chickens. The results showed that the antigen contents of the six batches of virus liquid were 1×10^7.7-1×10^8.1 TCID₅₀/0.1 mL,and the results of the two kinds of cells on the same virus sample were consistent; and there was no difference between repeated determinations. The neutralizing antibody titers of IBDV-positive serum samples were 8-17 lb; there was no difference in the neutralizing antibody detection results of the same serum sample between the two kinds of cells. The results suggested that DF-1 cells could replace CEF cells for antigen contents determination of IBDV fluid and be used as the culture substrate for determination of serum neutralizing antibody titer.