Abstract: OBJECTIVE To investigate the surveillance of clinical microbial cultures and ESKAPE (an acronym for the first letters of six types of bacteria: Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa and Enterobacter spp.) pathogens in a three-A hospital in Zhengzhou from 2015 to 2024, and to evaluate the effectiveness of the hospital's etiological testing and management of multidrug-resistant organisms. METHODS The hospital infection surveillance system was used to extract the microbial culture submission rate within the first 5 days before the use of therapeutic antimicrobial agents at People's Hospital of Zhengzhou. WHONET 5.6 was applied to analyze the isolation rates, specimen distribution and drug resistance of ESKAPE pathogens in patients at the hospital from 2015 to 2024. SPSS 22.0 software was used for trend analysis. RESULTS From 2015 to 2024, the microbial culture submission rate at the hospital increased from 15.99% to 31.28%, showing a continuous upward trend (P<0.001). The positive rate of specimens decreased (P<0.001). The most frequently detected pathogens were K. pneumoniae and E. coli, with the main specimen sources being sputum and urine. The proportion of sputum specimens showed a downward trend, while the proportions of bronchoalveolar lavage fluid and sterile specimens showed an upward trend, with statistically significant differences (P<0.001). From 2015 to 2024, the detection rate of carbapenem-resistant A. baumannii (CRAB) continuously decreased, while the detection rate of vancomycin-resistant E. faecium (VR-Efm) continuously increased. The detection rate of carbapenem-resistant P. aeruginosa (CRPA) decreased from 2015 to 2022 and increased from 2022 to 2024. The detection rate of carbapenem-resistant K. pneumoniae (CRKP) rapidly increased from 2015 to 2019, gradually decreased from 2020 to 2023, and showed a significant increase in 2024 compared to 2023, with all differences being statistically significant (P<0.001). The detection rates of methicillin-resistant S. aureus (MRSA) and carbapenem-resistant E. coli (CRCE) showed no significant trend. CONCLUSIONS The submission rate of microbial cultures and sterile specimens in this hospital continues to rise, while the detection rate of some ESKAPE drug-resistant organisms continues to decline, indicating that the hospital has achieved certain success in etiological testing and management of multidrug-resistant organisms. However, the detection rate of VR-Efm continues to rise, and the detection rates of CRPA and CRKP show an upward trend after initial decline, suggesting that the drug resistance situation in the hospital remains severe and requires attention.