To elucidate the putative role of human papillomavirus (HPV) infection in the etiology of esophageal cancer, 121 formalin-fixed, paraffin-embedded specimens originating from a non-high-incidence area for this carcinoma, from Slovenia, were screened for HPV infection using eight different polymerase chain reactions (PCR). Three different HPV consensus primer sets and four primer sets specific for HPV types 6, 16, and 18 failed to detect HPV DNA sequences in any of the tumor samples. Fragments of human beta-globin gene that served as internal controls were successfully amplified from 120 of 121 specimens. Our study confirms the opinion that most esophageal cancers originating from non-high-incidence geographic areas of this cancer are not associated with HPV infection. According to the studies reviewed, it is likely that HPV infection plays a much more significant role in esophageal carcinogenesis in those areas of the world with a high incidence of ESCC.