Objective: To determine the prevalence of herpes s implex virus (HSV) in malignant laryngeal lesions. Study Design: Retrospective review. Materials and Methods: Paraffin-embedded, histologically confirmed sp cimens containing benign laryngeal lesions, quamous cell carcinoma of the larynx, and squamous cell carcinoma of the oral cavity were identifi d from archived surgical specimens. Biopsies of normal-opp aring oral cavity tissue were also obtain d from fresh-frozen cadaver . These tissues were analyzed for the pre ence of HSV DNA using polymera se chain reaction techniques. Patient charts were revie wed for patient demographics, risk factors, tage, clinical course, treatment, and outcome. R esults: HSV was detected in nine laryngeal squamous cell carcinomas (75%) and in none of the benign laryngeal lesions (P = .0001). HSV was also found in three oral cavity squamous cell carcinomas (25%) and in none of the controls (P = .049). Conclusion: HSV is mor prevalent in squamous cell carcinoma of the larynx and oral cavity than in their respective control group , sugge ting a role for carcinogenesis. HSV is mor prevalent in squamous cell carcinoma of the larynx than of the oral cavity. Further studies to determine the role of HSV as a cocarcinogen and its inte rrelationship with other environmental factors in laryngeal cancer are warranted.