The present invention discloses Lactobacillus crispatus 262-1 and an inoculum comprising such strain and application thereof. The Lactobacillus crispatus 262-1 is a new strain of Lactobacillus genus and is preserved in the China General Microbiological Culture Collection Center with a preservation number of CGMCC No. 6469. The Lactobacillus crispatus 262-1 particularly is the dominant bacteria in the vagina, has good acid and H2O2 production capability, good adhesion to the vaginal epithelial cells and significant resistance to bacterial vaginosis and various vaginal infections, and is characterized by safety, non-toxicity, good stability and long term preservation. The present invention also relates to the use of the Lactobacillus crispatus 262-1 in the preparation of pharmaceuticals for preventing and/or treating gynecological diseases and its application in feminine care products such as medical instruments, disinfect products or cosmetics.