Recent studies identified viral and bacterial factors, including HSV-1 andH. pylori, as possible factors associated with diseases such as chronic tonsillitis and cancers, including head and neck squamous cell carcinoma (HNSCC). We assessed the prevalence of HSV-1/2 andH. pyloriin patients with HNSCC, chronic tonsillitis, and healthy individuals using PCR after DNA isolation. Associations were sought between the presence of HSV-1,H. pylori,and clinicopathological and demographic characteristics and stimulant use. HSV-1 andH. pyloriwere most frequently identified in controls (HSV-1: 12.5% andH. pylori: 6.3%). There were 7 (7.8%) and 8 (8.6%) patients with positive HSV-1 in HNSCC and chronic tonsillitis patients, respectively, while the prevalence ofH. pyloriwas 0/90 (0%) and 3/93 (3.2%), respectively. More cases of HSV-1 were observed in older individuals in the control group. All positive HSV-1 cases in the HNSCC group were associated with advanced tumor stage (T3/T4). The prevalence of HSV-1 andH. pyloriwas highest in the controls compared to HNSCC and chronic tonsillitis patients, which indicates that the pathogens were not risk factors. However, since all positive HSV-1 cases in the HNSCC group were observed only in patients with advanced tumor stage, we suggested a possible link between HSV-1 and tumor progression. Further follow-up of the study groups is planned.