AbstractThe majority of cervical cancer cases and associated deaths occur in low‐ and middle‐income countries (LMICs), where sociocultural barriers, poor access to prevention and care, and technical and practical difficulties hinder screening coverage improvement. Using urine specimens for human papillomaviruses (HPV) molecular screening through automated testing platforms can help to overcome these problems. We evaluated the high‐risk (HR) HPV detection performance of the Xpert® HPV test on GeneXpert® System (Cepheid), on fresh and dried urine (Dried Urine Spot [DUS]) samples as compared to an in‐house polymerase chain reaction (PCR) genotyping assay. Forty‐five concentrated urine samples collected from women with known cytological and HPV infection status, determined through in‐house PCR and genotyping assays, were tested “as is” and as DUS with the Xpert® HPV test. This system detected HR‐HPV in 86.4% of fresh and in 77.3% of dried urine samples collected from HPV+ women, correctly identifying HR‐HPV infection in 100% of women with low‐ and high‐grade lesions. High concordance (91.4%, k = 0.82) was found between PCR test and Xpert® HPV Test from urine. Urine‐based Xpert® HPV test seems to be a suitable screening test for detection of HR‐HPV infections associated with low‐ and high‐grade lesions requiring follow‐up monitoring or treatment. This methodology, relying on noninvasively collected samples and on available rapid testing platforms, could facilitate large, at‐scale screening programs, particularly in LMICs and rural areas, thus reducing adverse outcomes of HPV infection and facilitating achievement of the WHO cervical cancer elimination goal.