Objective To analyze the effect of caspofungin on inhibiting the formation of Enterococcus faecalis(E. faecalis) biofilm. Methods One strain of clinically isolated E. faecalis with strong positive biofilm and 10 strains of E. faecalis with different biofilm forming ability were selected, caspofungin intervention group and control group were set up at the same time, intervention group used caspofungin with the concentration of 20 μmol/L, control group used normal saline. Growth of planktonic strains and biofilm formation of E. faecalis in two groups were detected by crystal violet staining and microplate reader. Results Caspofungin had no inhibitory effect on the growth of planktonic clinical E. faecalis strains, but can significantly inhibit the formation of biofilm. Conclusion Caspofungin has a significant inhibitory effect on biofilm formation of E. faecalis.