Objective To investigate the distribution and drug resistance of pathogens isolated from blood culture in elderly patients in a hospital in Wuxi City, and provide evidence for rational empiric antimicrobial therapy for bloodstream infection(BSI). Methods Blood culture result and clinical data of outpatients, emergency and hospita-lized patients aged ≥ 65 years in a hospital from 2012 to 2016 were collected, WHONET 5.4 software was used to analyze the data. Results A total of 734 strains of pathogens were isolated from 15 102 blood specimens, 56.13%, 37.74%, and 6.13% of which were gram-negative bacteria, gram-positive bacteria, and fungi respectively. The top three species of pathogens were Escherichia coli, Klebsiella pneumoniae, and Staphylococcus epidermidis, accounting for 20.16%, 10.90%, and 7.90% respectively; positive strains of blood culture mainly came from intensive care unit (n=187, 25.48%). Resistance rates of Escherichia coli and Klebsiella pneumoniae to imipenem were 0 and 7.50% respectively, to amikacin were both<5%; resistance rates of Acinetobacter baumannii to commonly used antimicrobial agents were ≥ 50%, resistance rate to imipenem was 69.05%. Isolation rate of methicillin-resis-tant Staphylococcus aureus (MRSA) was 34.38%, which was lower than that of methicillin-resistant Staphylococcus epidermidis and methicillin-resistant Staphylococcus hemolyticus (91.38% and 100.00% respectively), no Staphylococcus spp. were found to be resistant to vancomycin and linezolid, resistance rates of Enterococcus faecium to vancomycin and linezolid were 3.03% and 6.06% respectively. Conclusion Gram-negative bacteria are the main pathogens isolated from blood culture of elderly patients, Escherichia coli and Klebsiella pneumoniae are the dominant pathogens, which are sensitive to imipenem and amikacin. Isolation rate of methicillin-resistant coagulase-negative Staphylococcus (MRCNS) is higher than that of MRSA, resistance rates of Enterococcus faecium to linezolid is higher than resistance rates to vancomycin.