Little is known about the prevalence and persistence of human papillomavirus (HPV) in older women, or about the association between hormone replacement therapy (HRT) use and HPV detection. Like oral contraceptives, HRT hormones may upregulate viral expression and subsequent risk of genital cancer. Postmenopausal women seeking routine gynecologic care were evaluated for HPV infection, testing cervical/vaginal cytology by PCR/DNA sequencing. The prevalence of HPV was 14%; 6% had oncogenic types and 5.8% had persistent infection. Although risk of HPV detection was non-significantly elevated after adjustment for age and HPV-related risk factors among current (adjusted odds ratio (OR)=2.3) and past (adjusted OR=3.2) hormone users compared to never users, past users had a significantly higher risk using combination HRT regimens with increasing duration (adjusted OR=1.8 per year; 95% confidence interval (CI)=1.1-3.1). These findings suggest that a significant percentage of older women are infected with HPV. This may put them at increased risk of genital cancers with longer use of HRTs. Additional studies are needed.