Detection and species identification of four human herpesviruses using polymerase chain reaction coupled with restriction endonuclease analysis.
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.
Demkin
Vladimir V
VV
Institute of Molecular Genetics, Russian Academy of Sciences, Moscow, Russia. vdemkin@img.ras.ru
Kruglova
Anna I
AI
Nikolaeva
Nadezhda P
NP
Yurchenko
Julia V
JV
D000818:Animals; D002460:Cell Line; D003587:Cytomegalovirus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D017931:DNA Primers; D004279:DNA, Viral / Q000032:analysis; D015252:Deoxyribonucleases, Type II Site-Specific; D006561:Herpes Simplex / Q000175:diagnosis; D018259:Herpesvirus 1, Human / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D018258:Herpesvirus 2, Human / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D004854:Herpesvirus 4, Human / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
D017931:DNA Primers; D004279:DNA, Viral; D015252:Deoxyribonucleases, Type II Site-Specific; C040158:GTAC-specific type II deoxyribonucleases; C027046:TCGA-specific type II deoxyribonucleases