Abstract: 　Objective　To investigate the distribution and antibiotic resistance of isolated pathogens in neonatal sepsis. Methods The results of blood culture and drug susceptibility test in neonates sepsis from January 2012 to June 2013 were retrospectively analyzed. Results One hundred and thirty-two strains were detected in the blood samples, with 100 （75.76%） Gram-positive bacteria, 30 (22.73%) Gram-negative bacteria and 2 (1.52%) fungus. Staphylococcus epidermidis, Escherichia coli and Staphylococcus aureus were the three most common pathogens. Gram-positive cocci was strongly resistant to penicillin (100.00%), erythromycin, selectrin and ampicillin/sulbactam (62.50%-100.00%), but still sensitive to vancomycin and teicoplanin. The resistance rate of Gram-negative bacilli to ampicillin was 100.00%, and the resistance rate to cefatriaxone, selectrin and cefuroxime was 61.54%-100.00%. The resistance rate to imipenem and piperacillin/tazobactam was lower. Conclusions The selection of sensitive antibiotics should be based on the pathogens and drug resistance testing for the treatment of neonatal sepsis.