The early detection of herpes simplex encephalitis (HSE) represents a major problem in viral diagnostics. We have now established a test system based on PCR for the specific amplification of HSV-DNA in cerebrospinal fluid (CSF) samples followed by oligonucleotide hybridization. The test proved to be very sensitive. Five molecules of HSV 1 DNA still yielded positive signals after hybridization. The assay was applied to CSF samples from 6 patients with confirmed HSE. All except one CSF sample obtained from the 2nd to the 8th day after onset of neurological symptoms yielded positive results. The primers used also exhibit a certain degree of crossreactivity with HSV 2, as revealed by testing of a CSF sample from an HSV 2-infected child. No positive signals were obtained with human DNA and with DNA from CMV- and VZV-infected fibroblasts. Also 42 CSF samples of patients suffering from other diseases of the CNS yielded negative results. The present results indicate that the detection of HSV DNA in CSF by PCR represents a valuable tool for the early diagnosis of HSE.