Objective To understand the colonization of Staphylococcus aureus (SA) and other cultivable symbiotic bacteria in nasal cavity, and provide reference for the formulation of prevention and control measures of infection and healthcare-associated infection (HAI) caused by opportunistic pathogenic bacteria (especially SA) colonized in nasal cavity. Methods Nasal vestibule specimens from military medical university healthy students who studied clinical medicine course in a hospital were collected and inoculated on various kinds of bacterial culture media by microbial automation system for bacterial culture and colony observation, the isolated strains were identified by mass spectrometry, SA was performed antimicrobial susceptibility testing. Results A total of 161 nasal vestibular specimens were collected, 549 bacterial strains of nasal colonization were isolated, including 493 strains of gram-positive and 56 strains of gram-negative bacteria, the top three bacteria were Corynebacterium accolens (151 strains), Staphylococcus epidermidis (150 strains) and Corynebacterium pseudodiphtheriticum (54 strains).There were 72 kinds of bacteria carrying patterns, 21 (13.04%) students were detected Staphylococcus aureus, Staphylococcus epidermidis and Corynebacterium accolens simultaneously, which was the most common patterns. Colonization rates of nasal SA and methicillin-resistant Staphylococcus aureus were 25.46% (41/161) and 4.35% (7/161) respectively, resistance rates of SA to penicillin, erythromycin and clindamycin were 39.02%-78.05%. Conclusion Nasal colonized bacteria in these university students are mainly normal flora, it is necessary to pay attention to the possibility of self-infection and HAI caused by colonized SA and other opportunistic pathogens.