Forty carbapenem-resistant Klebsiella pneumoniae isolates were recovered from 28 patients from various sites in an intensive care unit in Zhejiang Provincial People's Hospital, China, over a 6 month period. PFGE analysis indicated that the 40 strains were all closely related. The MICs of carbapenems varied from 16 to >256 µg ml⁻¹. Conjugation studies with Escherichia coli resulted in the transfer of reduced carbapenem susceptibility from the original isolates. All K. pneumoniae and E. coli transconjugants produced K. pneumoniae carbapenemase 2 (KPC-2), and most of them produced TEM, SHV and CTX-M. Additionally, 27 isolates and 27 E. coli transconjugants carried the qnr gene (25 were qnrB2 and 2 were qnrS1). K. pneumoniae harboured several plasmids, and bla(KPC-2) was located on a 55 kb plasmid. SDS-PAGE and ompK35/36 gene sequence analysis of OMPs suggested that porins in K. pneumoniae are expressed normally. The MICs of the carbapenems did not change in the presence of CCCP. Thus, production of KPC-2 appears to play an important role in resistance to carbapenems, although other mechanisms may be involved. The bla(KPC-2) gene is associated with several antibiotic-resistance genes, such as bla(TEM), bla(SHV), bla(CTX-M) and qnr.