Species identification of all eight human herpesviruses with a single nested PCR assay

There are eight currently known human herpesviruses, all of which are capable of latent persistence and reactivation following primary infection. Herpesvirus induced disease is common, widespread, and associated with significant morbidity, particularly in the immunocompromised human host. Current methods of herpesvirus detection include viral culture and polymerase chain reaction (PCR). A robust PCR method based upon amplification of the highly conserved herpesvirus DNA polymerase gene that is capable of detection of all eight human herpesviruses, including EBV and HHV-6 subtypes, from clinical material is described. Species identification of PCR products is accomplished by either of two methods, chemiluminescent dot blot hybridization and heteroduplex mobility shift assay, both of which allow for simultaneous detection of multiple herpesviruses. This method should prove useful for rapid and accurate species identification of all eight human herpesviruses from clinical material.