Background: The most common sexually transmitted infection is infection by human papilloma virus (HPV). Although the main transmission route of HPV is through sexual intercourse, the contact of the skin with the genital area skin is another known method for transmission. This study aims to examine the awareness and practice of female beauty salon staff about HPV and its transmission in pubic hair removal using wax. Materials and Methods: This cross-sectional study was conducted on 200 female beauty salon staff who do bikini waxing in Shiraz, Iran. Demographic information, awareness, and practice of them were gathered using the researcher-constructed questionnaire. The statistical analysis was performed using the SPSS software version 18. Results: Mean awareness and practice scores of beauty salon staff about HPV and its transmission in bikini waxing were 26.6 ± 12.8 and 5.5 ± 0.8, respectively, and in average range. The results showed that there was a significant relationship between practice and education (P = 0.04), but the relationship between the awareness and education was not seen (P = 0.72). There was no statistically significant relationship between age and work experience with awareness and practice of staff (P > 0.05). Conclusion: The awareness and practice of beauty salon staff about HPV and its transmission in bikini waxing are moderate. Therefore, it seems that the implementation of continuous educational program by the Union of women's beauty salons is necessary to increase and improve the awareness and practice of female beauty salon staff.