Objective To investigate the distribution and antimicrobial resistance difference of Klebsiella pneumoniae (KP) causing healthcare-associated infection (HAI) and community-associated infection (CAI) in a tertiary first-class hospital for three consecutive years, and provide evidence for rational use of antimicrobial agents in clinical practice. Methods KP isolated from hospitalized patients with CAI and HAI between November 2016 and October 2019 were retrospectively analyzed, constituent ratio and antimicrobial resistance difference of KP in different departments and different specimen sources were compared, change trend of antimicrobial resistance was analyzed. Results A total of 1 104 strains of KP were isolated, including 410 strains in HAI group and 694 in CAI group. KP in both groups were mainly isolated from sputum specimens, and the main department was intensive care unit. Except that the resistance rates of KP to tigecycline were both 0 in two groups, resistance rates to other antimicrobial agents in HAI group were all higher than those in CAI group (all P<0.05). Antimicrobial resistance rates of KP in CAI group were all <30%, resistance rates of KP to ceftriaxone and compound sulfamethoxazole in HAI group were more than 50%, resistance rate to imipenem was 18.54%. By trend χ² test, resistance rates of KP to ceftria-xone, cefepime, cefoxitin, cefoperazone/sulbactam, imipenem, amikacin, tobramycin, levofloxacin, and ciprofloxacin in HAI group all showed an upward trend (all P<0.05); resistance rates of KP to piperacillin/tazobactam, cefoxitin, cefoperazone/sulbactam, imipenem, and ciprofloxacin showed an upward trend in CAI group (all P<0.05), while resistance rate to gentamicin showed a downward trend (P=0.004). Conclusion Antimicrobial resis-tance rates of KP in HAI are higher than those in CAI, resistance rates to multiple antimicrobial agents in HAI group increase year by year, KP HAI and CAI should be treated differently.