Objective To investigate the changes of multidrug-resistant organisms (MDROs) in the First People’s Hospital of Longquanyi District of Chengdu around its overall relocation. Methods The First People’s Hospital of Longquanyi District of Chengdu was overall relocated on December 31st, 2016. The detection rates of MDROs and the changes in nosocomial infections before the relocation (from 2015 to 2016) and after the relocation (from 2017 to 2020) were retrospectively analyzed. Results A total of 83634 qualified specimens were submitted for inspection, 8945 strains of pathogenic bacteria were detected, and the detection rate of pathogenic bacteria was 10.70%, showing an increasing trend in yearly detection rates of pathogenic bacteria (χ2trend=8.722, P=0.003); among them, 1551 MDRO strains were detected, and the detection rate of MDROs was 17.34%, showing an increasing trend in yearly detection rates of MDROs (χ2trend=11.140, P=0.001). The detection rate of pathogenic bacteria before relocation was lower than that after relocation, and the difference was statistically significant (9.64% vs. 11.08%; χ2=35.408, P<0.001); there was no significant difference in the detection rate of MDROs before and after relocation (16.32% vs. 17.66%; χ2=2.050, P=0.152). From 2015 to 2020, the detection rates of pathogenic bacteria from sputum+throat swab specimens (χ2trend=81.764, P<0.001) and secretion+pus specimens (χ2trend=56.311, P<0.001) showed increasing trends, while the detection rates of pathogenic bacteria from blood specimens (χ2trend=110.400, P<0.001), urine specimens (χ2trend=11.919, P=0.001), and sterile body fluid specimens (χ2trend=20.158, P<0.001) showed decreasing trends. The MDRO detection rates of Escherichia coli (χ2trend=21.742, P<0.001), Staphylococcus aureus (χ2trend=47.049, P<0.001), and Pseudomonas aeruginosa (χ2trend=66.625, P<0.001) showed increasing trends, while the MDRO detection rates of Klebsiella pneumoniae (χ2trend=2.929, P=0.087) and Acinetobacter baumannii (χ2trend=0.498, P=0.481) showed no statistically linear trend, but the MDRO detection rate of Acinetobacter baumannii dropped significantly in 2017. In the targeted monitored MDROs, the proportions of nosocomial infections in methicillin-resistant Staphylococcus aureus (χ2trend=4.581, P=0.032), carbapenem-resistant Enterobacteriaceae (χ2trend=8.031, P=0.005), and carbapenem-resistant Pseudomonas aeruginosa (χ2trend=6.692, P=0.010) showed decreasing trends; there was no statistically linear trend in the proportion of nosocomial infections in carbapenem-resistant Acinetobacter baumannii (χ2trend=0.597, P=0.440); only one strain of vancomycin-resistant Enterococcus was detected in 2017, and no nosocomial infection occurred. Conclusions The overall detection rate of pathogenic bacteria and MDROs in this tertiary general hospital around relocation showed increasing trends year by year. The detection rate of pathogenic bacteria after relocation was higher than that before relocation, but the detection rate of MDROs after relocation did not differ from that before relocation. The proportion of nosocomial infections among the targeted monitored MDROs decreased.