Human herpesvirus type 6 variants identified by single-strand conformation polymorphism analysis.
Six human herpesvirus 6 (HHV-6) variants were analyzed for heterogeneity using the polymerase chain reaction (PCR) and single-strand conformation polymorphism (SSCP). Two independent DNA regions were selected: a fragment of the gene U11 (position 18966-21578) coding for a basic phosphoprotein, the major antigenic structural protein pp100; and a fragment from an open reading frame (ORF) area of the gene U67, previously referred to as 13R (position 102458-103519), coding for a product of unknown function. The two PCR systems based on the above DNA sequences yielded products of 187 bp and 223 bp, respectively. DNA obtained from three laboratory reference strains (U1102, R104 and St.W.) and from HHV-6 infected peripheral white blood cells of bone marrow transplant patients and blood donors was used to test the applicability of two different SSCP analysis systems for the identification of HHV-6 variants using amplicons derived by PCR from the two genomic regions described above (U11 [pp100], U67 [13R-ORF]). The generation of characteristic SSCP patterns enables the rapid differentiation of HHV-6 A and B strains for the classification of variants derived from clinical samples, reducing the need for expensive and time-consuming direct sequencing analyses.
Virchow-Klinikum, Abteilung für Innere Medizin mit Schwerpunkt Hämatologie und Onkologie, Humboldt Universität, Berlin, Germany.
F
F
Wilborn
T
T
Binder
F
F
Neipel
R
R
Blasczyk
W
W
Siegert
D001483:Base Sequence; D001773:Blood Cells; D002460:Cell Line; D004262:DNA Restriction Enzymes; D004279:DNA, Viral / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D014644:Genetic Variation; D015654:Herpesvirus 6, Human / Q000145:classification / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D018807:Polymorphism, Single-Stranded Conformational; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
D004279:DNA, Viral; D004262:DNA Restriction Enzymes