ObjectiveTo investigate the distribution and antimicrobial resistance of major pathogens causing bloodstream infection in a tertiary hospital, so as to guide the clinical rational use of antimicrobial agents.MethodsBlood specimens from patients with suspected bloodstream infection in this hospital between January 2012 and January 2015 were cultured, identified and performed antimicrobial susceptibility testing, the major pathogens and antimicrobial susceptibility testing results were analyzed statistically.ResultsA total of 1 658 strains of pathogens were isolated from 19 326 blood culture specimens, isolation rate was 8.58%, in which gramnegative bacilli, grampositive cocci, and fungi accounted for 58.57% (n=971), 39.38% (n=653), and 2.05% (n=34) respectively, the major gramnegative bacillus, grampositive coccus, and fungus were Escherichia coli, coagulasenegative staphylococci (CNS), and Candida albicans respectively. The isolated pathogens mainly distributed in departments of hematology, gastroenterology, and cardiovascular surgery. Detection rates of extendedspectrum βlactamases (ESBLs)producing Escherichia coli and Klebsiella pneumoniae were 56.35% and 36.14% respectively, but were highly sensitive to cefoperazone / sulbactam and carbapenems (resistance rates <3%). The percentage of methicillinresistant coagulasenegative staphylococcus (MRCNS) and methicillinresistant Staphylococcus aureus (MRSA) were 78.60％ and 32.28％respectively, but were highly sensitive to vancomycin and linezolid(resistance rates<1%).ConclusionEscherichia coli and CNS are predominant pathogens causing bloodstream infection in this hospital, antimicrobial resistance rates are high; regular analysis on distribution and antimicrobial resistance of pathogens causing bloodstream infection is helpful for the prevention and control of infection caused by multidrugresistant organisms.