Herpesviruses (especially cytomegalovirus [CMV] and Epstein-Barr virus [EBV]) have been implicated in the pathogenesis of chronic (Rasmussen) encephalitis associated with epilepsy. To assess the presence of herpesvirus genes in brain tissue from epileptic children with chronic (usually Rasmussen type) encephalitis, DNA was extracted from surgically resected brain tissue and studied by the polymerase chain reaction using primers specific for CMV, varicella zoster virus, herpes simplex virus, EBV, and human herpesvirus 6 genes. By this technique evidence for the presence of low levels of CMV and EBV genes was detected in most brain specimens from encephalitis patients and in several brain specimens from patients without encephalitis (eg, cortical dysplasia, gliosis, or encephalomalacia) who also had intractable epilepsy. Occasionally, both EBV and CMV genes were found in the same brain. Signal strength for both CMV and EBV was much lower in epileptic brains than in the brains of acquired immunodeficiency syndrome patients with CMV encephalitis or brain lymphoma. We found evidence for infection of the brain by human herpesvirus 6 in only one patient with encephalitis. Polymerase chain reaction technology applied to resected brain tissue from epileptic patients may provide evidence for or against viral-mediated pathogenesis of Rasmussen encephalitis or other types of encephalitis. The small amounts of EBV and CMV genes found suggest that herpesvirus infection of the brain does not directly cause Rasmussen encephalitis. © 1993.