Objective To study the distribution and drug resistance of pathogens in neonatal septicemia in order to provide clinical guidance for antibiotic usage.Methods This retrospective study analyzed blood culture and clinical data from 83 confirmed neonatal septicemia patients and the blood collection cultures were analyzed.Results A total of 84 strains were isolated from 83 cases of blood specimens,Gram positive bacteria,Gram negative bacteria and fungi were 38(45.2%),41(48.8%),5(6.0%),respectively.Gram positive bacteria was mainly coagulase negative staphylococcus and staphylococcus aureus,which were 13(15.5%) and 8(9.5%) respectively.Gram negative bacteria was mainly Escherichia coli and Klebsiella pneumonia,which were 25(29.8%) and 9(10.7%) respectively.Gram positive bacteria were found high resistance to penicillin G,amoxicillin clavulanate potassium,oxacillin and clindamycin,from 34.2% to 73.7%,but they were sensitive to vancomycin,teicoplanin and linezolid.Gram negative bacteria were found high resistance to ampicillin(82.9%),the constituent ratio of the extended spectrum βlactamases(ESBLs) was 34.1%,carbapenem resistant strains was not found.All fungi were sensitive to azoles.Conclusion Gram negative bacteria are the major pathogens in neonatal septicemia,with high infection rate of Escherichia coli and high constituent ratio of the ESBLs,and antimicrobial agents should be chosen according to blood culture and antimicrobial susceptibility results.