ObjectiveTo investigate the clinical distribution and antimicrobial resistance of Streptococcus agalactiae(S. agalactiae) in neonatal intensive care unit(NICU), and provide reference for antimicrobial use and intervention measures. MethodsSpecimens from neonates in the NICU of a hospital in 2010-2014 were collected, the department sources and antimicrobial susceptibility testing results of 62 strains of S. agalactiae isolated from children were analyzed. Results62 strains of S. agalactiae were mainly distributed at fullterm NICU, accounting for 64.52%; the main source of specimens was blood, accounting for 90.33%, followed, by cerebrospinal fluid (6.45%）, sputum, and secretion（both were 1.61%）. S. agalactiae had the highest resistance rate to tetracycline(79.03%)；resistance rates to erythromycin and clindamycin were both 74.19%, resistance rate to levofloxacin was 40.32%， susceptibility rates to penicillin and ampicillin were both 100%. ConclusionS. agalactiae infection mainly occurred in neonates in fullterm NICU, and has high resistance rate to multiple antimicrobial agents, penicillin and ampicillin can be used as the preferred antimicrobial agents for the treatment of S. agalactiae infection.