The invention discloses a method for detecting acute toxicity of rare earth tailing pond surrounding groundwater pollution by using freshwater luminescent bacteria. The freshwater luminescent bacteria are Vibrio qinghaiensis Q67; the method comprises steps as follows: a to-be-detected sample is treated and diluted, and the pH value is adjusted; an activated Vibrio qinghaiensis Q67 solution is added to the to-be-detected sample for a reaction; meanwhile, a diluent for dilution replaces the to-be-detected sample to serve as a contrast; the luminescent intensity of the to-be-detected sample and the luminescent intensity of the contrast are detected by a bioluminescence tester; the result is calculated, standard reference poison ZnSO4 is set, and the toxicity effect is judged according to the result. The method is rapid, sensitive, accurate and convenient to popularize and apply. Meanwhile, Vibrio qinghaiensis, the freshwater luminescent bacteria, is used as a test strain, and the influence caused by salinity required by marine luminescent bacteria on luminescence is avoided. The reference poison is ZnSO4, and the influence on the ecological environment and human health is reduced.