Abstract: OBJECTIVE To establish the environmental bacteria library of pharmaceutical microbial testing laboratory. METHODS 248 strains of bacteria and 6 strains of fungi were collected from the pharmaceutical microbiology testing laboratory from different parts, equipments and materials and the activities of personnel through 7 months working, and those bacteria were identified by biochemical identification method with Vitek2 Compact. Those 90 strains of bacteria without exact results of biochemical identification and 6 strains of fungi were identified by 3500 MicroSeq Gene Sequencing Analysis System for bacterial 16S rDNA and fungal LSU rDNA sequencing, and 6 strains of Staphylococcus aureus of a sampling collection were analyzed of homology with Diversilab system. RESULTS The environmental bacteria library was initially established by sorting and analyzing of the identification results of the collected 254 strains of microorganisms. CONCLUSION A technology platform is provided to evaluate the risk of pollution and to investigate the adverse events in the future. Technical guidance is also provided to the pharmaceutical companies which are establishing the data base of environmental microorganisms and tracing of microbial contamination.