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BACKGROUND This paper is to explore the composition and antibiotic resistance of pathogens isolated from blood cultures of patients with bloodstream infection (BSI) in the intensive care unit (ICU). METHODS A retrospective study has been performed to identify the pathogens isolated from blood cultures of 147 patients with BSI in the ICU of our Hospital over 12 years (January 2008 to December 2019). The antibiotic resistance and susceptibility of the isolates were tested. RESULTS Among the 152 pathogens isolated from positive blood cultures, 61.84% were Gram-positive bacteria, 34.87% were Gram-negative bacteria, and 3.29% were fungi. The most frequently isolated pathogens were coagulase-negative Staphylococci, Staphylococcus aureus, and Escherichia coli. The pathogen distributions remained stable over 12 years. Gram-positive cocci were highly susceptible to vancomycin, linezolid, and teicoplanin. Extended-spectrum β-lactamase-producing pathogens were highly susceptible to amikacin, imipenem, meropenem, and polymyxin B. Acinetobacter were relatively sensitive to amikacin, trimethoprim-sulfamethoxazole, and polymyxin, whereas Pseudomonas aeruginosa were sensitive to amikacin, gentamicin, levofloxacin, imipenem, meropenem, and ciprofloxacin, with resistance rates less than 30%. CONCLUSIONS Gram-positive bacteria were the dominant pathogens isolated from blood cultures of ICU-BSI patients and were susceptible to vancomycin, linezolid, and teicoplanin. The antibiotic susceptibility of Gram-negative bacteria varies greatly. Our results provide important information to guide the treatment decisions in the management of ICU-BSIs.