Objective To investigate the distribution characteristics and antimicrobial resistance of common Enterobacterales in a hospital, and provide reference for clinical rational use of antimicrobial agents. Methods All non-repetitive strains of Enterobacterales isolated from clinical specimens in a hospital from 2010 to 2019 were collected. Detection, clinical distribution, specimen distribution and antimicrobial resistance of different kinds of bacteria were analyzed. Results A total of 19 384 strains of Enterobacterales were collected, 45.32% (8 784 strains) were Escherichia coli, 25.11% (4 867 strains) were Klebsiella pneumoniae, 5.67% (1 099 strains) were Enterobacter cloacae, 3.67% (711 strains) were Serratia marcescens, 3.29% (638 strains) were Klebsiella oxytoca, and 16.95% (3 285 strains) were other Enterobacterales. Most strains of Escherichia coli were isolated from urine (42.53%), followed by respiratory tract (21.63%) and blood (16.33%); Klebsiella pneumoniae was mainly isolated from respiratory tract (69.37%), followed by urine (9.33%) and blood (8.73%). Escherichia coli was mainly isolated from department of urology surgery (29.16%), followed by pediatrics (8.86%); Klebsiella pneumoniae was mainly isolated from department of neurosurgery (22.52%), followed by pediatrics (13.71%). Resistance rates of Enterobacterales to ampicillin, piperacillin and cefazolin were higher in 2018-2019, and resistance rates to imipenem and meropenem increased significantly, isolation rate of carbapenem-resistant Enterobacterales (CRE) increased year by year, with statistical significance (P < 0.05). Conclusion Antimicrobial resistance of Enterobacterales is not optimistic, clinicians should take effective measures to prevent the emergence of CRE, and it is also very important to prevent and control cross infection in hospital.