       Document 0443
 DOCN  M9610443
 TI    Quantitative PCR for human herpesviruses 6 and 7.
 DT    9601
 AU    Secchiero P; Zella D; Crowley RW; Gallo RC; Lusso P; Laboratory of Tumor
       Cell Biology, National Cancer Institute,; Bethesda, Maryland 20892-4255,
       USA.
 SO    J Clin Microbiol. 1995 Aug;33(8):2124-30. Unique Identifier : AIDSLINE
       MED/96057598
 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.
 DE    Acquired Immunodeficiency Syndrome/COMPLICATIONS/VIROLOGY  Base Sequence
       CD4-Positive T-Lymphocytes/VIROLOGY  DNA Primers/GENETICS  DNA,
       Viral/GENETICS  Evaluation Studies  Herpesviridae
       Infections/COMPLICATIONS/DIAGNOSIS  Herpesvirus 6,
       Human/*GENETICS/ISOLATION & PURIF/PATHOGENICITY  Herpesvirus 7,
       Human/*GENETICS/ISOLATION & PURIF/PATHOGENICITY  Hodgkin's
       Disease/COMPLICATIONS/VIROLOGY  Human  Molecular Sequence Data
       Polymerase Chain Reaction/*METHODS/STANDARDS  Reference Standards
       Support, Non-U.S. Gov't  JOURNAL ARTICLE

       SOURCE: National Library of Medicine.  NOTICE: This material may be
       protected by Copyright Law (Title 17, U.S.Code).

