The presence of human papillomavirus (HPV) DNA in esophageal brushings from human immunodeficiency virus (HIV)-seropositive hosts was investigated in a cross-sectional study. Oral and esophageal brushings from individuals scheduled for esophagogastroscopy (53 HIV-positive and 61 age-matched HIV-negative patients) were tested for the presence of HPV DNA by a consensus L1 polymerase chain reaction assay. HPV DNA was detected in esophageal brushings of 9 (17%) of the 53 HIV-seropositive patients and 0 of the 61 HIV-negative individuals. HPV-16 DNA was the most frequently detected. No proliferative mucosal lesion was noted in individuals with HPV-positive esophageal brushings. Cytological smears were done for 6 of the 9 patients with HPV-positive esophageal brushings, and epithelial atypia was recorded for 1. HIV infection and a history of genital herpes were strong independent predictors of HPV, suggesting that HPV is transmitted sexually in the esophagus.