Aim: Human papillomavirus (HPV) detection is very important for the evaluation of prevention strategies in cervical cancer and other HPV-related diseases. This study aimed to assess the prevalence of high-risk HPV infection in a cohort of Qatari residents, and correlate this with cytology and potential risk factors. Method: The study utilized cervical cytology and HPV DNA testing methods, high-risk screen real-time PCR, to detect high-risk HPV genotype infections in a sample of Qatar’s female population. A series of 95 women attending the Gynecology/Oncology Clinic at Hamad Medical Corporation between August 2007 and May 2008 were enrolled in the study. Cervical smears isolated from patients were subjected to real-time PCR to confirm the presence of HPV DNA. The smears were characterized as atypical squamous cells of undetermined significance (ASCUS); low-grade squamous intraepithelial lesions (LGSIL) and high-grade squamous intraepithelial lesions (HGSIL). As a secondary outcome, risk factors for HPV infection were also assessed. However, the samples were not case-controlled for these. Results: A total of 28 women had normal smears and 65 had abnormal smears. Among the 65 women in the study population with abnormal smears, 33 had ASCUS (18 of them were HPV DNA positive), 21 had LGSIL (18 were HPV DNA positive), eight had HGSIL (four were HPV DNA positive) and the remaining three had squamous cell carcinoma (two were HPV DNA positive). Conclusion: The overall prevalence of high-risk HPV in our study population (n = 95) was 64%. The relatively high prevalence of HPV 52, 56 and 16 among the study group has important implications in vaccine prophylaxis in Qatar.