Objective To analyze the distribution and antimicrobial resistance of clinically isolated bacteria in the intensive care units (ICUs) of Gansu Province, and provide basis for the rational antimicrobial use for ICU patients in this region. Methods Antimicrobial resistance monitoring data of clinically isolated bacteria from ICUs in hospitals of antimicrobial resistance surveillance system in Gansu Province from January 2019 to December 2022 were collected. Antimicrobial resistance were detected by disk diffusion method, minimum inhibitory concentration (MIC) method and E-test method, and the test results were judged according to the standards of American Clinical and Laboratory Standards Institute (CLSI). Data were analyzed using WHONET 5.6 and SPSS 26.0 softwares. Results From 2019 to 2022, Gram-negative bacteria were the main clinical isolates in ICUs in Gansu Province, with detection rates of 74.5%, 75.0%, 75.9%, and 75.5%, respectively. Klebsiella pneumoniae had the highest detection rate. Among Gram-positive bacteria, Staphylococcus aureus had the highest detection rate, all around 25%. The detection rates of methicillin-resistant Staphylococcus aureus were 43.9%, 43.3%, 41.8%, and 42.7%, respectively. The detection rates of methicillin-resistant coagulase negative Staphylococcus (MRCNS) were 83.9%, 82.4%, 81.8%, and 77.5%, respectively. Detection rate of Enterococcus faecalis showed a downward trend (χ²=5.229, P=0.021). The resistance rate of Pseudomonas aeruginosa to multiple antimicrobial agents was less than 33.9%, and the resistance rates to aminoglycosides and quinolones showed a decreasing trend (P < 0.05). The resistance rates of Acinetobacter baumannii to various antimicrobial agents such as aminoglycosides and carbapenems presented a rising trend (P < 0.05). The detection rates of MRCNS and carbapenem-resistant Klebsiella pneumo-niae showed a decreasing trend (both P < 0.05). Conclusion From 2019 to 2022, the main clinically isolated pathogenic bacteria in ICUs in Gansu Province were Gram-negative bacteria. The detection rate of multidrug-resistant organisms has increased and drug resistance was severe, which should be paid attention in clinic. It is necessary to strengthen pathogen monitoring and antimicrobial resistance analysis, use rational antimicrobial agents, as well as strengthen infection prevention and control measures, so as to avoid the emergence of more antimicrobial-resistant bacteria due to healthcare-associated infection and cross transmission of antimicrobial-resistant organisms.