Aims To prevent the spread of brucellosis, a simple and rapid vertical flow technology (RVFT) for the detection of antibodies targeting brucellosis was developed. Methods and Results In this study, Brucella sp. lipopolysaccharide was purified and used to detect brucellosis antibodies. Sheep IgG was used as a negative control. Colloidal gold-labeled recombinant staphylococcus aureus protein A was sprayed on a fibreglass membrane to prepare immunogold pads. Rapid vertical flow technology was used to detect Brucella in 1668 Sheep, 2743 bovine, 674 red deer and 420 human samples. The results indicated that the accuracy of this assay can reach 98%. Conclusions The established RVFT uses a single multifunctional buffer that can be used to detect antibodies in serum, plasma, whole blood and other biological samples while preserving the advantages of lateral-flow immunoassays. Significance and Impact of the Study This technology would be of great use in primary medical units and veterinary stations, and it is of great significance for the control of epidemic diseases.