Comprehensive PCR-based assay for detection and species identification of human herpesviruses.
The description and evaluation of a PCR-based assay for the detection and species identification of the eight known human herpesviruses are presented. Two primer pairs targeting well-conserved regions of the genome allowed the amplification of the DNAs of all known human herpesviruses at a high level of sensitivity (10 to 100 genome copies for most viruses). Identification of the virus species was achieved through restriction enzyme digestion with BamHI and BstUI, which yielded fragment sizes that were characteristic for each herpesvirus. Furthermore, it was demonstrated that this restriction enzyme panel allowed the discrimination between human herpesvirus 6 variant A and variant B. This assay format was validated over the course of 1 year in a clinical virology laboratory setting, where it was shown that it readily detected human herpesviruses, including occasional multiple infections, in a variety of clinical samples. The PCR assay was compared to isolation and electron microscopy for the detection of herpes simplex (HSV) and varicella-zoster virus (VZV) in clinical samples. All specimens positive by conventional methods were also positive by PCR. However, in a number of clinical specimens in which HSV or VZV could not be detected by conventional methods, PCR was able to demonstrate the presence of the virus.
Johnson
G
G
Department of Paediatric Laboratory Medicine, The Hospital for Sick Children, Toronto, Ontario, Canada.
Nelson
S
S
Petric
M
M
Tellier
R
R
D001769:Blood / Q000821:virology; D001826:Body Fluids / Q000821:virology; D002555:Cerebrospinal Fluid / Q000821:virology; D015245:Deoxyribonuclease BamHI / Q000378:metabolism; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism; D006564:Herpesviridae / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D006566:Herpesviridae Infections / Q000175:diagnosis* / Q000821:virology*; D006801:Humans; D008854:Microscopy, Electron; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D015183:Restriction Mapping; D012680:Sensitivity and Specificity; D012867:Skin / Q000821:virology; D014776:Virus Cultivation
D015245:Deoxyribonuclease BamHI; C039566:CGCG-specific type II deoxyribonucleases; D015252:Deoxyribonucleases, Type II Site-Specific