Objective: To analyze detection rate and changing trend of clinical multi-drug resistant bacteria(MDRB) in recent 5 years, and explore drug resistance characteristics of carbapenem resistant gram-negative bacteria, so as to provide reference for nosocomial infection prevention and control and empirical treatment of antibiotics in clinical practice. Methods: Seven kinds of MDRB isolated from clinical specimens of hospitalized patients during January 2017 to December 2021 were retrospectively analyzed, including methicillin-resistant Staphylococcus aureus (MRSA), vancomycin resistant Enterococcus (VRE), extended spectrum β-lactamases producing Escherichia coli (ESBL-eco), extended spectrum β-lactamases producing Klebsiella pneumoniae (ESBL-kpn), carbapenem resistant Enterobacteriaceae (CRE), carbapenem resistant Acinetobacter baumannii (CR-AB) and carbapenem resistant Pseudomonas aeruginosa (CR-PA). The rate of carbapenem resistant gram-negative bacilli to common antibiotics and the carbapenemase production of CRE strain were analyzed with SPSS 20.0. Results: VRE was not detected in this study. Except ESBL-eco, the isolated rates of the other five MDRB increased in the past five years. The detection rates of MRSA, ESBL-kpn, CRE, CR-AB and CR-PA fluctuated between 37.0%-53.6%, 19.3%-34.3%, 2.5%-10.6%, 34.1%-63.4% and 11.3%-22.8%, respectively. It revealed that 65.4% CRE, 72.0% CR-AB, and 59.3% CR-PA isolates were isolated from ICU. The resistant rates of CRE and CR-AB to quinolones, cephalosporins and β-Lactamase inhibitor complex were 90.0%. Among 272 CRE strains, 221 strains were Klebsiella pneumoniae, accounting for 81.3%. The carbapenemase was detected in 90 strains of CRE, in which three kinds of carbapenemases (KPC, NDM and IMP) were detected and the constituent ratio of KPC enzyme was 90.0%. Conclusions: The isolated rates of six MDRB in this study are much lower than the average level in Shanghai and national tertiary hospitals in the same period, but the isolated rates of MRSA, ESBL-kpn, CRE, CR-AB and CR-PA show an upward trend, which should be paid attention to, especially the carbapenem resistant gram-negative bacteria in ICU. In this study, 90 strains of CRE tested produced three kinds of carbapenem enzymes, in which KPC enzymes showed a high proportion.