Human papillomavirus (HPV) has been established as a causative agent in the development of oral squamous cell carcinoma (OSCC). Specifically, HPV types 16 and 18 are known to be prevalent in oral cancers. This cross-sectional study aimed to determine the prevalence of HPV types 16 and 18 in OSCC cases in Qazvin province, Iran. Thirty-eight paraffin-embedded samples of OSCC were selected, and DNA extraction was performed using the Roche High Pure FFPE DNA isolation kit. The quality of the extracted DNA was assessed through PCR amplification of the human β-Globin gene. The HPV detection was carried out using SYBR green-based real-time PCR with GP5+ and GP6+ primers targeting the L1 region of HPV. The HPV genotyping was conducted on positive samples using specific primers. Statistical analysis was performed between HPV infection in OSCC and age, sex, and anatomical location. This study analyzed 38 biopsy specimens obtained from male and female OSCC patients, with an average age of 64 years. Among these samples, 13 tested positive for HPV, resulting in a prevalence rate of 34.2%. The age group with the highest HPV infection rate was 61-70 (10.5%) years. Notably, HPV type 16 was detected in 21.0% of samples, HPV type 18 in 10.5%, and other viral subtypes in 2.6%. No statistically significant correlation was found between HPV prevalence and gender or age. The findings indicated that 34.2% of OSCC samples in the Qazvin province harbor HPV, with types 16 and 18 being the most common in tumors affecting the tongue. Additionally, no association was observed between HPV infection and age or gender. To address HPV as a risk factor for OSCC, public health initiatives such as vaccination, awareness campaigns, and accessible healthcare services should be implemented. They are, furthermore, incorporating HPV DNA testing into practice.