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The aim of this study was to determine whether an association exists between Epstein Barr Virus (EBV) and esophageal squamous cell carcinoma (ESCC) among Iranian patients. The incidence of the cancer in Iranians is high. EBV has been implicated in a number of human malignancies, many of epithelial origin. The etiology of ESCC appears to involve genetic and environmental factors. Tumerous and histologically normal paraffin-embedded esophageal tissue of 30 patients, and 6 cancerous and 4 non-cancerous freshly frozen tissue specimens of ESCC patients were studied. The samples were evaluated for the presence of EBV by PCR and Southern blot hybridization. The EBV genome was detected in only one non-cancerous specimen. It was concluded that EBV is probably not associated with ESCC among Iranian patients. This is consistent with the results of studies on several but not all other populations. The implications of the geographical variation are discussed.