Background: As one of the most prevalent cancers in men, prostate cancer is a condition with multiple causes. Viral infections have been identified as one of the major sources of elevated incidence of prostate cancer. Objectives: The purpose of this research was to assess the association of the risk of prostate cancer and its aggressiveness with seropositivity of herpes simplex virus 2 (HSV-2) and/or human HERPESVIRUS 8 (HHV-8). Methods: Totally, 103 men with prostate cancer as cases and 81 healthy individuals as controls were included in this case-control analysis and provided a serum sample. The specific IgG antibodies against HSV-2 and HHV-8 were screened by enzyme-linked immunosorbent assay (ELISA). To determine the association between HSV-2, HHV-8, prostate-specific antigen (PSA) level, and demographic variables with incidence of prostate cancer, univariate and multivariate logistic regression models were applied. Results: The results of the univariate logistic regression model showed a statistically significant association between HSV-2 and HHV-8 seropositivity, PSA level, age, and smoking with prostate cancer incidence (P 0. 20). The multivariate logistic regression model results after adjusting for the potential confounding variables showed a significant statistical association between the mean of PSA level [adjusted odds ratio (OR): 3. 44; 95% CI: 2. 15-5. 51; P < 0. 001) and incidence of prostate cancer. Moreover, the results of univariate and multivariate logistic regression model showed a significant statistical association between age [adjusted OR: 0. 88; 95% confidence interval (CI): 0. 81-0. 95; P = 0. 001] and HSV-2 and also significant statistical association was found between PSA (adjusted OR: 1. 02; 95% CI: 1. 005-1. 03; P = 0. 006) and HHV-8. Conclusions: Although the seroprevalence of HSV-2 and HHV-8 was higher in patients with prostate cancer than in the control group, it cannot be concluded that there is a significant association between the seropositivity of these viruses and prostate cancer incidence. However, the findings showed a significant statistical association between age and seropositivity of HSV-2 and also a significant statistical association between PSA levels and seropositivity of HHV-8.