Background: Herpes simplex viruses have been implicated as a cause of cardiovascular disease (CVD) due to their widespread distribution and ability to infect human vascular endothelial cells. Objectives: The present study aimed to evaluate the link between Herpes simplex infections and cardiovascular disease. Methods: This case-control study involved 236 patients aged 35 - 65, 118 with known cardiovascular disease and 118 controls. Patients’ cardiovascular disease evaluation was based on data from questionnaires, a specialist’s physical examinations, and paraclinical tests. Herpes simplex viruses (HSV)-antibodies (IgM, IgG) were determined using ELISA in serum samples, and the biochemical blood indicators were analyzed to examine their relationship to the level of HSV antibodies. All the data were analyzed using SPSS software version 20. Results: IgM antibodies against herpes simplex were negative among all 236 patients. The positivity rate of IgG antibodies against herpes simplex was 58.8 and 51.2% in the case and control groups, respectively with no significant difference (P = 0.253). Patients with cardiovascular disease had a 0.587-fold higher positive rate of IgG antibodies than patients in the control group (odds ratio (OR) = 0.587). The mean age of patients with positive IgG antibodies was significantly lower than patients with negative IgG antibodies (P = 0.012). IgG positivity among men and women (P = 0.670) and different job statuses (P = 0.866) were not significantly different. Conclusions: The positivity rate of IgG antibodies against herpes simplex was not significantly different among the study groups. Although patients with positive IgG antibodies’ mean age was lower than those with negative IgG antibodies, the gender and job status were not different.