PMID- 7559960
OWN - NLM
STAT- MEDLINE
DCOM- 19951122
LR  - 20210526
IS  - 0095-1137 (Print)
IS  - 1098-660X (Electronic)
IS  - 0095-1137 (Linking)
VI  - 33
IP  - 8
DP  - 1995 Aug
TI  - Quantitative PCR for human herpesviruses 6 and 7.
PG  - 2124-30
AB  - A quantitative PCR assay for the detection of human herpesvirus 6 (HHV-6) 
      (variants A and B) and HHV-7 DNAs in clinical samples was developed. The assay 
      uses a nonhomologous internal standard (IS) for each virus that is coamplified 
      with the wild-type target sequence in the same vial and with the same pair of 
      primers. This method allows for a correction of the variability of efficiency of 
      the PCR technique. A standard curve is constructed for each experiment by 
      coamplification of known quantities of the cloned HHV-6 or HHV-7 target templates 
      with the respective IS. Absolute quantitation of the test samples is then 
      achieved by determining the viral target/IS ratio of the hybridization signals of 
      the amplification products and plotting this value against the standard curve. 
      Using this assay, we quantitated the amount of HHV-6 or HHV-7 DNA in infected 
      cell cultures and demonstrated an inhibitory effect of phosphonoformic acid on 
      the replication of HHV-6 and HHV-7 in vitro. As the first clinical application of 
      this procedure, we performed preliminary measurements of the loads of HHV-6 and 
      HHV-7 in lymph nodes from patients with Hodgkin's disease and AIDS. Application 
      of this quantitative PCR method should be helpful for elucidating the pathogenic 
      roles of HHV-6 and HHV-7.
FAU - Secchiero, P
AU  - Secchiero P
AD  - Laboratory of Tumor Cell Biology, National Cancer Institute, Bethesda, Maryland 
      20892-4255, USA.
FAU - Zella, D
AU  - Zella D
FAU - Crowley, R W
AU  - Crowley RW
FAU - Gallo, R C
AU  - Gallo RC
FAU - Lusso, P
AU  - Lusso P
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - J Clin Microbiol
JT  - Journal of clinical microbiology
JID - 7505564
RN  - 0 (DNA Primers)
RN  - 0 (DNA, Viral)
SB  - IM
MH  - Acquired Immunodeficiency Syndrome/complications/virology
MH  - Base Sequence
MH  - CD4-Positive T-Lymphocytes/virology
MH  - DNA Primers/genetics
MH  - DNA, Viral/genetics
MH  - Evaluation Studies as Topic
MH  - Herpesviridae Infections/complications/diagnosis
MH  - Herpesvirus 6, Human/*genetics/isolation & purification/pathogenicity
MH  - Herpesvirus 7, Human/*genetics/isolation & purification/pathogenicity
MH  - Hodgkin Disease/complications/virology
MH  - Humans
MH  - Molecular Sequence Data
MH  - Polymerase Chain Reaction/*methods/standards
MH  - Reference Standards
PMC - PMC228347
EDAT- 1995/08/01 00:00
MHDA- 1995/08/01 00:01
PMCR- 1995/08/01
CRDT- 1995/08/01 00:00
PHST- 1995/08/01 00:00 [pubmed]
PHST- 1995/08/01 00:01 [medline]
PHST- 1995/08/01 00:00 [entrez]
PHST- 1995/08/01 00:00 [pmc-release]
AID - 10.1128/jcm.33.8.2124-2130.1995 [doi]
PST - ppublish
SO  - J Clin Microbiol. 1995 Aug;33(8):2124-30. doi: 10.1128/jcm.33.8.2124-2130.1995.