〔Abstract〕 Objective To analyze the relationship between carbapenem-resistant Klebsiella pneumoniae virulence factor, capsule serotype and biofilm formation. Methods Clinical strains of Klebsiella pneumoniae in Huazhong University of Science and Technology Union Hospital from 2011 to 2016 were collected. The minimum inhibitory concentration (MIC) against meropenem and imipenem was determined by microbroth dilution method to select carbapenem-resistant clinical strains.The polymerase chain reaction (PCR) technology was used to amplify virulence genes (rmpA、rmpA2、wabG、uge、fimH、mrkD、ycf、wcaG、 entB、iron、hly、cnf、aerobactin) and pod serotypes (K1、K2、K5、K20、K54、K57). The biofilm forming ability of Klebsiella pneumoniae was detected by crystal violet staining. Results A total of 41 carbapenem-resistant strains were screened. The average OD₅₅₀ value of Klebsiella pneumoniae after crystal violet staining was 1.07 ± 0.37. There was no statistical significance between Klebsiella pneumoniae capsule serotype and biofilm formation. The results of single factor analysis and multiple linear regression analysis of virulence factors showed that wabG and iron virulence factors were related to the formation of Klebsiella pneumoniae biofilm. Conclusion Carbapenem-resistant Klebsiella pneumoniae was more likely to form biofilms with wabG and iron virulence factors.