The objective of this study was to evaluate the presence of Staphylococcus aureus in the nasal cavity of children attending a daycare center in the town of Francisco Beltrão-PR, by comparing the frequency between age groups and genders. Antimicrobial susceptibility testing with different drugs was also carried out in the bacterial isolates. A prospective study involving 200 children aged 2 and 4 years, grouped by gender, was carried out. Samples were collected from the nasal vestibules of children, incubated and identified. Antimicrobial susceptibility testing was carried out in 30% of the S. aureus colonies. The results showed that 35% of children (10% female and 25% male) were colonized by S. aureus. The frequency of colonization in both male age groups was similar, with 201.93 ± 13.10 CFU/plate for 2-year-old and 266 ± 12.60 CFU/plate for 4-year-old. However, colonization averages for females were different, with 30.43 ± 1.17 CFU/plate for 2-year-old and 394.38 ± 10.70 CFU/plate for 4-year-old. In the antimicrobial susceptibility testing, some strains were resistant to oxacillin, erythromycin and tetracycline, in addition to showing intermediate susceptibility to cephalothin.