Objective To analyze carbapenemase production and molecular epidemiological characteristics of clinically isolated carbapenem-resistant Klebsiella pneumoniae (CRKP) from two hospitals in Wuhan City. Methods 42 non-repetitive CRKP strains were collected from two hospitals in Wuhan City from January to October 2018, carbapenemase production was screened by Carba NP test, carriage of carbapenemase genes was detected by polyme-rase chain reaction (PCR), horizontal transfer of drug resistance genes was analyzed by plasmid conjugation test, phylogenetic relationship was analyzed by pulsed-field gel electrophoresis (PFGE). Results Of 42 CRKP strains, 14 were positive for Carba NP test, 10 of which were amplified NDM-1 gene and 3 amplified KPC-2 gene. A total of 13 CRKP strains were positive for carbapenem gene detection, 12 of which were successful in plasmid conjugation test. PFGE showed that CRKP strains carrying NDM-1 gene were classified into 6 types, there was no dominant type; CRKP strains carrying KPC-2 gene were of the same type. Conclusion NDM-1 and KPC-2-producing CRKP strains were detected, plasmid conjugation test suggested that plasmid-mediated horizontal transfer might play an important role in the dissemination of carbapenemase genes.