Abstract: A two-step dot immunogold filtration assay(T-DIGFA) was established to detect antibodies against schistosome in livestock. In this method, 1 μL lixivium from blood paper for examination was dotted on a nitrocellulose membrane(1.3 cm×1.3 cm) in a plastic box with a hole(Φ0.6 cm) on it’s face center, 50s～90 s later, 100 μL extract of schistosome egg labelled with colloidal gold were added on nitrocellulose membrane in the hole. If there were anti-schistosome antibodies in blood paper lixivium, a red mark would appear. If there were no anti-schistosome antibodies in the lixivium, no red mark could be seen.In this study 139 blood paper positive samples from cattle with schistosomiasis diagnosed by feces hatching miracidium method and 130 blood paper negative samples from cattle without schistosomiasis were detected with T-DIGFA, the results showed that schistosomiasis positive coincidence rate was 100% and negative coincidence rate was 99.2% compared with feces hatching miracidium method. The detected result was according to that using HRP-SPA-Dot-ELISA method completely. T-DIGFA could also detect the antibodies against schistosome in lixivium of blood paper from cattle and rabbits infected with schistosome cercaria artificially after 7 days (including 7 days) and from the cattle infected by one twain of schistosome naturally. There was no cross reaction with the antibodies of the Faciola hepatica, Trypanosoma evansi and ascariasis. The same 125 samples of blood paper were detected by five operators with five colloidal gold prepared on different time, respectively, no different results were found between operators or between various batches of colloidal gold. T-DIGFA reagent kit that were stored in 4-8 ℃ for 6 months or kept in room temperature for 3 months still kept valid. The tests verified that T-DIGFA method have high sensitivity, specificity and stability. It is suitable for fast diagnosis and general survey for schistosmiasis of animals.