Objective To rapidly screen Staphylococcus aureus (SA) colonized in nasal cavity by using eSwab flocked nylon swab transport system and chromogenic culture medium, understand SA colonization in outpatients in department of otorhinolaryngology-head and neck surgery (ORL-HNS), and provide basis for prevention and control of SA infection as well as healthcare-associated infection(HAI). Methods eSwab was used to collect nasal vestibule specimens of patients in department of ORL-HNS in a hospital, specimens were inoculated on blood agar medium and methicillin-resistant SA(MRSA)/SA chromogenic culture medium by WASPLab microbial automated system, SA colonies were photographed and observed at 16 and 40 hours respectively, results were verified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOF MS), antimicrobial susceptibility testing and mecA gene detection. Results A total of 200 nasal vestibule specimens were collected, 48 strains of SA were isolated, 23(47.9%) of which were methicillin-sensitive SA(MSSA), and 25(52.1%) were MRSA; nasal colonization rate of SA and MRSA were 24.0% and 12.5% respectively. The average reporting time of SA positive screening was (17.6±6.1) hours. Coincidence rate of screening culture with MALDI-TOF MS identification and drug-resistance phenotype detection were both 100.0%, coincidence rate with mecA gene detection was 97.9%. Conclusion The rapid screening method based on eSwab flocked nylon swab transport system combined with chromogenic culture medium has higher accuracy and shorter reporting time, which can be used for rapid screening of SA colonization.