High risk forms of the human papilloma virus (HPV) are generally accepted as necessary causative agentsfor cervical cancer. Recently, a possible relation between HPV and oral squamous cell carcinoma (OSCC) hasalso been noticed. The present study was conducted to investigate the prevalence of HPV infection in OSCCs inWuhan city. DNA samples were collected from fresh tissues in 200 patients with OSCC and 68 normal controls.The polymerase chain reaction and direct sequencing were used to identify the HPV types in the samples. Theprevalence of HPV of all types in the OSCC group was higher than in the control group (55/200 vs 2/68, OR=11.5,95% CI=2.6-50.2). HPV16 and HPV18 were the main types detected, with HPV6 was the only low-risk typeidentified. High-risk HPV types HPV16 and HPV18 are prevalent in OSCC patients and may participate in thedevelopment of OSCC with traditional risk factors, tobacco and alcohol, possibly exerting synergistic effects.The results of multinomial logistic regression showed that those who smoked, consumed alcohol and with HPVinfection have the highest risk of developing oral cancer (OR=13.3, 95% CI=3.1-56.8). Adjusted for age, smokingand alcohol use, HPV infection was independently associated with oral squamous cell carcinoma.