Abstract: Objective: To investigate the molecular epidemiological feature of clinical carbapenemase-producing Klebsiella pneumoniae isolates, and then provide the instruction for the prevention and control of nosocomial infections. Methods: Three clinical carbapenem-resistant Klebsiella pneumoniae isolates were collected from sputum and blood samples of newborns who were admitted into a hospital of Shandong province. Their identification and antibiotic susceptibility tests were performed with the Vitek 2 compact system. The production of carbapenemase was detected by the modified Hodge test. The resistance genes in these isolates, including the carbapenemase genes such as bla_KPC, bla_BIC, bla_OXA-48, bla_NDM, bla_IMP, bla_VIM, bla_SPM, bla_AIM, bla_GIM, bla_SIM and bla_DIM, β-lactamase genes such as bla_CTX-M, bla_SHV, bla_TEMand bla_OXA-1, AmpC β-lactamase genes such as bla_MOX, bla_CIT, bla_DHA, bla_ACC and bla_EBC, plasmid mediated quinolone resistance genes such as qnrA, qnrB, qnrS, oqxA and oqxB, and fosfomycin resistance gene such as fosA3, were detected by PCR, and confirmed by DNA sequencing. The homology of these isolates was assessed by pulsed field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). The transferability and incompatibility groups of plasmids were determined by the conjugation test, Southern blot hybridization and PCR based replicon typing (PBRT) method, respectively. Results: All of three Klebsiella pneumoniae isolates had positive Hodge test, and were highly resistant to most of tested antibiotics. They carried with bla_NDM-1, bla_TEM-1, fosA3, oqxA and oqxB genes. The three Klebsiella pneumoniae isolates had the same banding pattern in PFGE, and all belonged to ST22 grouping. The conjugation test and Southern hybridization analysis showed that the donor bacterium carried with plasmids with bla_NDM-1 (pTA-NDM, about 220 000 bp) could successfully transferred pTA-NDM into the recipient bacterium (E. coli J53AziR) and that the pTA-NDM belonged to IncA/C type plasmid. Conclusion: To our knowledge, it may be the first report that the New Delhi metallo beta-lactamase-1(NDM-1) producing Klebsiella pneumoniae ST22 isolates simultaneously carried with bla_NDM-1, bla_TEM-1, fosA3, oqxA and oqxB genes spread in a neonatal intensive care unit, which should be paid attention to by us.