Abstract: Objective To set up marine bacteria identification database by matrix assisted laser desorption ionization time-of-flight mass spectrometry,aiming to provide support for rapid identification of pathogens isolated from marine bacterial infections. Methods Totally 530 marine bacteria were collected by our research team from 6 sea areas and 158 sites from 2006 to 2016.The amplification and sequencing of the 16srDNA of bacteria were performed after isolating and culturing.The FlexControl software was used to detect the protein spectrum and the Biotyper software was used to construct and assess the database.In order to compare if there was difference between the correct identification rates of different databases,the bacterial identification was performed by the commercial,in-house and extended databases,respectively. Results The in-house marine bacteria database included 32 genus,82 species,and 421 strains spectrums in sum.The identification by 16srDNA sequencing was defined as golden standard.Among the validation strains,80 were identified by the commercial database with the accuracy rate of 80.8%,and 97 were identified by extended database (in-house+commercial) with the accuracy rate of 98.0%.The difference was significant (P<0.001). Conclusion Theidentification ability of in-house marine bacteria database is superior to the commercial one and is beneficial to the diagnosis and therapy to the marine bacterial infections.