A simple and rapid diagnostic assay system was developed for detecting and identifying human alpha-herpesviruses, such as herpes simplex virus types 1 and 2 and varicella-zoster virus, by polymerase chain reaction. This system was based on an amplification step, using primers that bind the DNA polymerase consensus sequence of alpha-human herpesviruses, and a detection step, using non-radioactive virus-specific probes. This method could be used to amplify any human alpha-herpesviruses, and each virus-specific probe was highly specific for identification of the amplified product. This system is readily applicable for implementation in the clinical laboratory.