To establish the characteristics of bacterial colonization and antibiotic resistance in the newborn in Neonatal Intensive Care Unit (NICU) of Beijing Obstetrics and Gynecology Hospital, Capital Medical University, pharyngeal swab specimens of hospitalized newborns in NICU were collected from August 1, 2014 to May 31, 2015, and subjected to bacterial culture and drug resistance analysis. A total of 677 swabs were included in this study and 34.0% were positive on culture detection. Among these positive cases, 159 (69.1%) were Gram-positive cocci, and 71 (30.9%) were Gram-negative bacilli. Streptococcus viridans, Kocuria kristinae and Staphylococcus epidermidis were the main colonized Gram-positive cocci. Escherichia coli, Acinetobacter baumannii and Klebsiella pneumoniae were the main colonized Gram-negative bacilli. Drug resistance of the isolates to commonly used antibiotics was studied and presented.