Human papillomavirus (HPV) is the most common virally sexually transmitted disease, and high-risk types of HPV have been implicated in over 90% of cervical cancers worldwide. Current preventative me measures of cervical cancer include routine Pap smear evaluation, which appears to be effective. Current preventative measures for cervical cancer include routine Pap smear evaluation, which appear to be effective. However, a small but significant proportion (>10%) of females in the United States have either never had a Pap smear or (>30%) do not have them on a routine basis. This may be due, in part, to the invasiveness and discomfort of the requirement pelvic examination. Although HPV cannot be routinely grown in the laboratory, its DNA can be deleted by amplification techniques such as PCR. Detection of HPV DNA from cervical swab or cervical lavage specimens has been used as an epidemiological tool to determine the prevalence rates of HPV infection. These procedures also require a pelvic examination that limits its widespread applicability. A method that is equally sensitive and efficient but does not require a pelvic examination to detect HPV infection will be able to identify more women at risk for cervical cancer and greatly aid in epidemiologic surveys. The recent advances in the diagnosis of gonorrhea and chlamydia infection by screening urine using amplification techniques demonstrate the feasibility of diagnosis a cervical infection by a urine test. Preliminary date have demonstrated the ability to detect HPV DNA in urine specimens from women at high risk for HPV infection. For these reasons, we hypothesize that a urine PCR test for the detection of HPV DNA will reflect the state of infection for the cervic. The goal of this proposal is to fully develop and validate a urine PCR test for HPV DNA detection that can be utilized for epidemiologic screening purposes. We propose to initially develop the urine PCR assay for HPV DNA detection by studying 20 women with no detectable HPV DNA in their urine. The ability to detect beta-globin DNA (internal control for the presence of cells) and known amounts of clon4ed HPV DNA spiked into these urine specimens will be measured and optimized. Next, urine will be obtained from 50 women previously tested to have HPV DNA detected in cervical/vaginal swabs. The extraction method by the initial experiments will be verified by testing these known HPV positive "field" specimens. Finally, utilizing the conditions optimized in specific aims #1 and verified in specific aim #2, a cohort of 250 women at high-risk and 250 women at low-risk for HPV infection will be enrolled. Paired urine and cervico-vaginal swabs will be obtained and the ability to detect any HPV DNA, any high-risk HPV DNA and type-specific HPV DNA will be compared. A validated urine test for HPV DNA detection could be used to better define the epidemiology of HPV, to explore the natural history of HPV infection, and to identify women at higher risk for cervical cancer.