Detection and species identification of four human herpesviruses using polymerase chain reaction coupled with restriction endonuclease analysis

Abstract

A polymerase chain reaction (PCR) based assay for detection and species identification of four human herpesviruses, including herpes simplex virus types 1 and 2, Epstein-Barr virus, and cytomegalovirus was developed. The detection of the herpesviruses was achieved by seminested PCR with three primers targeting well-conserved regions within the DNA-polymerase gene. Virus species were identified by simple restriction enzyme digestion of the amplified products with TaqI or RsaI. In comparison with mono-specific nested PCR assays the tetra-specific assay demonstrated similar specificity and sensitivity with reference and clinical samples. The tetra-specific assay is sensitive, cost effective, and can be used for examination of clinical samples of different origin.