Detection of human herpesvirus 6 by reverse transcription-PCR.
The role of human herpesvirus 6 (HHV-6) in disease beyond primary infection remains unclear. We have developed and validated a new reverse transcription-PCR (RT-PCR) assay for HHV-6 that can determine the presence of HHV-6 in clinical specimens and differentiate between latent and replicating virus. Peripheral blood mononuclear cells from 109 children were evaluated for HHV-6 by RT-PCR, DNA PCR, and viral culture. Of these samples, 106 were suitable for analysis. A total of 20 samples were positive for HHV-6 by culture and DNA PCR, of which 19 were positive by RT-PCR (sensitivity, 95%). All 28 samples from children that were negative by viral culture, but positive by DNA PCR, were negative for viral transcripts by our RT-PCR assay. One positive RT-PCR result was observed in 56 samples that were negative by tissue culture and DNA PCR. This indicates a low rate of false-positive results (1.2%) and a specificity of 98.8%. This RT-PCR assay can reliably differentiate between latent and actively replicating HHV-6 and should allow insight into the pathogenesis of this ubiquitous virus.
Norton
R A
RA
Department of Microbiology and Immunology, University of Rochester Medical Center, Rochester, New York 14642, USA. Renee_Norton@urmc.rochester.edu
Caserta
M T
MT
Hall
C B
CB
Schnabel
K
K
Hocknell
P
P
Dewhurst
S
S
D001483:Base Sequence; D002675:Child, Preschool; D017931:DNA Primers / Q000235:genetics; D004279:DNA, Viral / Q000032:analysis / Q000235:genetics; D003951:Diagnostic Errors; D005069:Evaluation Studies as Topic; D005260:Female; D006566:Herpesviridae Infections / Q000175:diagnosis / Q000821:virology*; D015654:Herpesvirus 6, Human / Q000235:genetics* / Q000302:isolation & purification* / Q000502:physiology; D006801:Humans; D007223:Infant; D008297:Male; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods* / Q000706:statistics & numerical data; D012680:Sensitivity and Specificity; D014773:Virology / Q000379:methods; D014776:Virus Cultivation / Q000379:methods; D014779:Virus Replication
D017931:DNA Primers; D004279:DNA, Viral