Abstract: OBJECTIVE To investigate the etiological characteristics of mixed bloodstream infection in hospitalized patients, analyze the drug resistance, and explore the related factors for prognosis so as to provide guidance for clinical treatment. METHODS The medical records were collected from 216 patients who had mixed bloodstream infection from Aug 2008 to Dec 2015, the results of microbiological examination were analyzed, and the drug resistance was observed by using Whonet5.6 software. RESULTS Of the patients who had the mixed bloodstream infection, 40.28% were in department of surgery, and 19.91% were isolated from department of hepatobiliary surgery. A total of 442 strains of pathogens were isolated, of which 25.11% (111 strains) were Enterococcus spp, 18.78% (83 strains) were Escherichia coli, 15.61% (69 strains) were Klebsiella pneumoniae. There were 45 pathogen combinations patterns, of which 6 mixed pathogen were dominant pattern. There was no carbapenem-resistant E.coli strains, and the E.coli strains were highly susceptible to amikacin and piperacillin-tazobactam; the drug resistance rate of the K.pneumoniae to imipenem was 10.14%; the drug resistance rate of the P.aeruginosa was less than 30.00%; the drug resistance rate of the Acinetobacter baumannii to most of the antibiotics was more than 80.00%; the drug resistance rate of the E. faecium was generally higher than that of the E. faecalis; the drug resistance rate of the E.faecium to vancomycin was 3.45%; and the vancomycin-resistant E.faecalis strains were not detected. CONCLUSION It is necessary to reasonably use antibiotics according to the drug resistance of the different combinations of pathogens causing the mixed bloodstream infection and strictly monitor the drug susceptibility at the same time.