Background: Prevalence and factors associated with sexual dysfunction in HIV-positive women are poorly known. Methods: This was a cross-sectional study in a cohort of HIV-infected women. Clinically stable women were invited to participate in a female sexual dysfunction (FSD) evaluation with Female Sexual Function Index (FSFI) exploring desire, arousal, lubrication, orgasm, pain and satisfaction. An FSFI score <23 was used for defining FSD. Variables evaluated included body appearance satisfaction, interference of body changes with habits, social life and attitudinal aspects of body image, health-related quality of life, hormonal assessment, menopause, cumulative exposure to antiretroviral drug classes and immune-virological parameters. Lipodystrophy was defined according to the HIV Outpatient Study definition. Results: A total of 185 women completed the FSFI. The mean (±sd) age was 42 years (±5), 27% had CDC stage C, the mean (±sd) CD4 + T-cell count was 508 cells/μl (±251) and median HIV RNA was 1.7 log 10 copies/ ml (interquartile range 1.7–2.6). Among 161 evaluable patients, 59 (32%) reported FSD. In a multiple linear regression analysis, desire, arousal and satisfaction domains were associated with interference of body changes with habits, social life and attitudinal aspects of body image (β=0.22, 95% confidence interval [CI] 0.06–0.37; β=0.29, 95% CI 0.10–0.48; and β=0.20, 95% CI 0.02–0.38, respectively). Lubrication and orgasm domains were associated with body image satisfaction (β=-0.49, 95% CI -0.88— 0.10 and β=-0.58, 95% CI -1.00—0.16, respectively). No significant associations with sex hormones, CDC stage, CD4 + T-cell count, HIV RNA viral load and cumulative exposure to antiretroviral drug classes were found. In women with FSD, severity of self-perceived abdominal fat accumulation showed a trend towards lower FSFI scores (ANOVA P=0.02). Conclusions: FSD was highly prevalent in this cohort. Self-perceived body changes was identified as its major determinant.