The aim of this study was to compare the acceptability of self-collected vs clinician-collected human papillomavirus (HPV) tests. Utilising a cross-sectional approach, 250 non-pregnant, 25-60-year-old women from Leon, Nicaragua self-collected vaginal swabs and brushes for HPV and received a pelvic examination for liquid-based cytology and reflex HPV. Subjects underwent colposcopy, with biopsy if indicated, and completed questionnaires comparing their perceptions of HPV testing methods. Acceptability indices were significantly higher for self-collected brushes (M = 18.40, SD = 2.73) and swabs (M = 18.48, SD = 2.41) than clinician-collection (M = 17.56, SD = 2.92), p < 0.001. Multiple regression analyses revealed that age, education, and reproductive history were not significant predictors of acceptability. A total of 81.1% of subjects reported they would self-collect in the future. Self-collected HPV tests are highly acceptable in terms of pain, comfort, privacy and embarrassment, and are thus a viable method of screening in low-resource areas.