The invention discloses a method for evaluating leaching toxicity of municipal waste incineration fly ash. The method comprises the following steps: S1, preparing a waste incineration fly ash sample;S2, preparing a leaching agent; S3, adding the sample prepared in the step S1 into the leaching agent prepared in the step S2 to prepare a leaching solution; S4, preparing a bacterial culture solution; S5, culturing the bacteria and evaluating the activity of the bacteria; S6, if the bacterial activity in the step S5 is qualified after being detected, further preparing a cell culture solution, andif the bacterial activity in the step S5 is unqualified after being detected, performing comet assay and western blot detection; S7, culturing cells; S8, taking the cells cultured in the step S7 formulti-cytotoxicity test, and evaluating the cell activity; S9, if the cell activity in the step S8 is detected to be qualified, ending the test, and if the cell activity is detected to be unqualified,performing comet assay; and S10, performing western blot analysis by the cells obtained in the step S7. According to the method, the toxic reaction of bacteria and biological cells to the leaching solution is directly utilized, and the method can be used for scientifically evaluating the toxicity detection of the waste incineration fly ash.