Objective To explore and analyze the characteristics and transmission routes of carbapenem-resistant Klebsiella pneumoniae (CRKP) strains in intensive care unit (ICU). Methods From January to October 2023, 17 clinical infection isolates (clinical infection group), 5 active screening isolates (active screening group), and 7 environmental isolates (environmental group) of CRKP in the liver surgery ICU of a hospital were selected and analyzed by whole-genome sequencing. The differences in resistance genes, virulence genes, and sequence typing (ST) were compared, and transmission routes were analyzed based on the phylogenetic tree. Results 29 strains of CRKP carried 4-18 resistance genes and 52-98 virulence genes, respectively. There were no statistically significant diffe-rences in genotype distribution of resistance genes, the number of virulence genes, and gene types among three groups of CRKP (all P>0.05). ST showed that 29 CRKP strains mainly consisted of two categories: ST11 and ST15. Based on the phylogenetic tree constructed from the core genome, there were 7 highly homologous groups of CRKP, among which 4 groups had clear epidemiological associations. Conclusion CRKP in ICU carries more resistance and virulence genes, and some strains are highly homologous in ST and phylogenetic tree, which may lead to cross transmission. In the future, prevention and control measures should be strengthened to reduce the transmission of CRKP.