Methods that used specimens from three genital sites (penile brushing [PB], urethral brushing [UB], and the retrieval of semen [SE]) from 50 men were examined for human papillomavirus (HPV) DNA detection. The rates of detection by PB, UB, SE, PB and UB, and PB and SE were 88.9%, 50.0%, 33.3%, 100%, and 97.2%, respectively. The use of PB and UB appears to be the most accurate method; as an alternative to UB, the use of SE with PB could be used to improve the rate of HPV DNA detection in men.Infection with high-risk human papillomavirus (HPV) is the main cause of cervical cancer (7). Since men have been suggested to play a likely role as reservoirs and vectors for HPV infection (8,12,28), screening of men may be relevant for the prevention of cervical cancer in women.In the absence of clinical lesions, the most reliable diagnostic strategy for men is testing for HPV DNA (25). As recently reviewed (10), HPV infection rates in males range from 1.0% to 82.9%. In addition to the different profiles of the patients and the different HPV assays used, this wide range of rates may also be due to the variation in the clinical material analyzed (e.g., penile surface, preputial cavity, glans, scrotum, urethra, semen, and urine) because of a lack of agreement on the anatomical sites that should be sampled. Of the few studies that compared different specimens from men (1, 2, 11, 22), some reported that penile brushing (PB) represents the most sensitive method, while others selected urethral brushing (UB) or the retrieval of semen (SE). To assess the most adequate procedure, useful information could be obtained by comparing different samples from "high-risk" men, such as the sexual partners of HPV-positive women (8,16,25). To this aim, this study researched specimens from different sites for HPV DNA by three different methods, namely, PB, UB, and SE. The specimens were from 50 partners (age range, 23 to 58 years; mean age, 36. Patients were instructed not to wash their genitalia on the day before the examination and to have 2 days of sexual abstinence. Three types of genital specimens were collected in parallel. The first type was collected by PB, which consisted of the collection of cells from the dorsal and the ventral surfaces of the penile shaft, sampled with a standard-sized, dry cottontipped swab, and cells from the inner part of the foreskin, coronal sulcus, frenulum, and glans, sampled with a salineprewetted cytobrush. Five to six backward and forward swab or brush movements were performed at each site, and all these samples were placed in the same tube containing 3 ml of phosphate-buffered saline (PBS). The second sample was obtained by UB, for which a very thin, saline-prewetted brush was inserted 1.5 cm into the urethra, rotated 360 degrees, and removed; when it was required by the patients, a 2% lidocaine gel was applied before sampling by UB. The cells obtained by UB were also placed in 3 ml of PBS. The third sample was obtained by SE, with the semen collected by masturbation and placed in sterile containers. If the...