Abstract: OBJECTIVE To explore the distribution of pathogenic bacteria and change of drug resistance in nosocomial bloodstream infection in 2014-2017. METHODS The data on blood samples of 1 485 patients who were admitted for examination in the hospital from Jan. 2014 to Jan. 2017 were retrospectively analyzed, including 588 cases of patients with blood infection. The distribution of pathogenic bacteria and change of drug resistance were analyzed according to the results of blood culture and drug susceptibility test. RESULTS From 588 samples of the 1 485 blood samples, 624 strains of pathogens were isolated and cultured, with an isolation rate of 39.60%. There were 361 strains of gram-negative bacteria accounting for 57.85%, 209 strains of gram-positive bacteria accounting for 33.49% and 54 strains of fungi accounting for 8.65%. The detection rate of gram-negative bacteria decreased from 2016-2017 to 2014-2015 (χ²=0.742, P=0.826), whereas the detection rate of gram-positive bacteria increased (χ²=0.428, P=0.916). The major gram-negative bacteria were sensitive to carbapenems, and had relatively high resistance rates to other antimicrobial drugs, with an increasing trend year by year. The major gram-positive bacteria showed no resistance to vancomycin, but rather high resistance rates to the commonly used antibiotics such as penicillin and ampicillin. The fungi isolated were sensitive to common antifungal agents. CONCLUSION The main pathogens of bloodstream infection are gram-negative bacteria, but the gram-positive bacteria show a trend of growth. The main pathogens have obvious resistance to common clinical antibacterial drugs, and show a trend of increasing year by year, which is worthy of clinical attention.