Pre- and non-occupational Post-Exposure Prophylaxis for prevention of HIV infection are recommended for adults at substantial risk for HIV infection. Women experiencing homelessness have increased risk for HIV infection compared with stably housed women. We conducted a survey of 74 sheltered women at Lotus House Women's Shelter (Lotus House) in Miami to assess risk behaviour as well as knowledge and perception of pre- and non-occupational post-exposure prophylaxis in this population. Of surveyed women, the 58.1% engaged in vaginal and/or anal sex while sheltered, and of sexually active women 55.4% reported inconsistent condom use; 83.8% of women reported no concern regarding HIV acquisition due to their behaviour. Few women surveyed (20.8%) had previously heard of pre- or non-occupational post-exposure prophylaxis. The majority (58.3%) of respondents indicated receptiveness to these prevention methods when introduced. Those indicating that they would consider pre- or non-occupational post-exposure prophylaxis were significantly younger than those indicating that they would not consider these prevention strategies (p = 0.004). Education and referral for pre- and non-occupational post-exposure prophylaxis should be considered for sheltered women at risk for HIV infection. Additional research to optimise implementation of biomedical prevention strategies in this population is needed.