Abstract Context: Oral squamous cell carcinoma (OSCC), the third most prevalent solid cancer in India with 45% incidence, is associated with 20%–50% of cases having human papillomavirus (HPV) infection; however, the current role of HPV in managing OSCC remains unclear, particularly in high-incidence areas such as Eastern India. Aim: The study aims to determine the prevalence of HPV in OSCC patients and evaluate its role as an independent risk factor in the progression of OSCC. Settings and Design: This cross-sectional study, conducted in an Eastern Indian tertiary care hospital, involved 140 identified and treated OSCC patients. Materials and Methods: Immunohistochemistry (IHC) was used to determine HPV immunoreactivity using the marker p16. The data were analyzed using SPSS version 27.0 with a significance level of P < 0.05. Results: Among 140 patients, 11 showed strong P16 positivity (>70%) for HPV, resulting in a prevalence rate of 7.86%. However, no statistically significant association of HPV (p16 positivity) with any groups was found. Interestingly, all P16-positive patients in our study had a history of tobacco use (9.5%), suggesting that HPV may not be considered an independent factor for oral cancer. Conclusion: In our cases, p16 positivity is seen in 7.85% of cases. This finding suggests that further sub-analysis concerning p16 as a prognostic marker and its therapeutic implication is further required in oral cancer. Correlation with PCR is also required to substantiate TNM staging of p16 by the IHC method.