Objective To analyze the resistance characteristics, main epidemic clones and biofilm formation ability of carbapenem-resistant Acinetobacter baumannii (CRAB) in a hospital, and provide reference for better prevention and control of CRAB infection. Methods 32 non-repetitive CRAB strains were collected from the hospital, antimicrobial susceptibility was detected by automatic microbiological analysis system, biofilm formation ability was detected by crystal violet staining, polymorphism of CRAB clones was analyzed by pulsed-field gel electrophoresis (PFGE). Results Resistance rate of 32 CRAB strains to tetracycline was the lowest (68.6%) and to other antimicrobial agents were all>78%. PFGE homology results showed that 32 CRAB strains could be divided into 9 different clone types (type A-I), each clone type consisted of 1-9 strains, the main prevalent clones were type A (n=9) and type E (n=7). Of 32 CRAB strains, 14 (43.8%) had biofilm formation ability and were weakly positive; there was no significant difference in biofilm formation ability among different clones (χ²=6.636, P=0.659). With the enhancement in biofilm formation ability, antimicrobial resistance rate increased in varying degree, but only gentamicin resistance rate between biofilm formation strains and biofilm non-formation strains was significantly different(χ²=4.879, P<0.05). Conclusion There are different clonal transmission of CRAB in this hospital, mainly type A and E, enhancement in biofilm formation ability can increase the drug resistance of strains.