ObjectiveTo investigate clinical distribution and antimicrobial resistance of Staphylococcus aureus（S. aureus）isolated from neonates,and provide guidance for rational use of antimicrobial agents.MethodsOne hundred and twelve strains of S. aureus isolated from neonates in Hunan Children's Hospital between January, 2008 and December, 2009 were analysed retrospectively.ResultsMost strains of S. aureus  were isolated from sputum and throat swabs in neonates with respiratory tract infection (65.18%,73/112）, the next were from blood, wound secretion and pus, which accounting for 12.50%, 12.50% and 7.14% respectively, resistant rate of S. aureus to clindamycin, erythromycin, amoxicillin and penicillin were high (41.08%~96.43%)； Methicillinresistant S. aureus (MRSA) were 20 strains (17.86%), drugresistance of methicillinsensitive S. aureus was significantly lower than that of MRSA (P=0.00 or P<0.05); All strains were sensitive to vacomycin and linezolid.ConclusionS.aureus is mainly isolated from respiratory tract specimens, and has strong resistance to antimicrobial agents, monitoring of pediatrics should be strengthened, and antimicrobial agents should be used rationally to prevent the emergence of drugresistant strains.