ObjectiveTo investigate the distribution and drug resistance of major pathogens in a neonatal intensive care unit (NICU), so as to provide evidence for the clinical therapy of infection.MethodsSpecies and antimicrobial susceptibility testing of the major pathogens isolated from NICU patients who were hospitalized between January 2011 and March 2012 were analyzed retrospectively.ResultsAmong 5 612 detected specimens, 3 060 (54.53%) pathogenic isolates were isolated. The major strains were Streptococcus viridans（29.93%）, Staphylococcus epidermidis（14.48%）, Moraxella catarrhalis（14.02%）, and Escherichia coli（9.48%）.The major grampositive cocci had very low resistant rates to vancomycin, teicoplanin and cefoperazone/sulbactam, the resistant rates were 0.00%-3.03%, 0.00%-5.83% and 6.73%-36.36%, respectively, while resistant rates to penicillin,erythromycin, ampicillin/sulbactam and cefoxitin were high (42.60%-100.00%). The major gramnegative bacilli had very low resistant rates to imipenem, piperacillin/tazobactam and cefoperazone/sulbactam,the resistant rates were 0.00%, 0.00%-9.05% and 0.00%-18.55%, respectively, while the resistant rates to cefuroxime, compound sulfamethoxazole and ampicillin/sulbactam were high（42.53%-100.00%）.ConclusionRealizing the distribution and drug resistance is helpful for the rational use of antimicrobial agents in clinic.