Transcriptomal analysis of varicella-zoster virus infection using long oligonucleotide-based microarrays.
Varicella-zoster virus (VZV) is a human herpes virus that causes varicella as a primary infection and herpes zoster following reactivation of the virus from a latent state in trigeminal and spinal ganglia. In order to study the global pattern of VZV gene transcription, VZV microarrays using 75-base oligomers to 71 VZV open reading frames (ORFs) were designed and validated. The long-oligonucleotide approach maximizes the stringency of detection and polarity of gene expression. To optimize sensitivity, microarrays were hybridized to target RNA and the extent of hybridization measured using resonance light scattering. Microarray data were normalized to a subset of invariant ranked host-encoded positive-control genes and the data subjected to robust formal statistical analysis. The programme of viral gene expression was determined for VZV (Dumas strain)-infected MeWo cells and SVG cells (an immortalized human astrocyte cell line) 72 h post-infection. Marked quantitative and qualitative differences in the viral transcriptome were observed between the two different cell types using the Dumas laboratory-adapted strain. Oligonucleotide-based VZV arrays have considerable promise as a valuable tool in the analysis of viral gene transcription during both lytic and latent infections, and the observed heterogeneity in the global pattern of viral gene transcription may also have diagnostic potential.
Kennedy
Peter G E
PGE
Glasgow University Department of Neurology, Southern General Hospital, Institute of Neurological Sciences, Glasgow G51 4TF, UK.
Grinfeld
Esther
E
Glasgow University Department of Neurology, Southern General Hospital, Institute of Neurological Sciences, Glasgow G51 4TF, UK.
Craigon
Marie
M
Scottish Centre for Genomic Technology and Informatics, Medical School, University of Edinburgh, Edinburgh EH16 4SB, UK.
Vierlinger
Klemens
K
Scottish Centre for Genomic Technology and Informatics, Medical School, University of Edinburgh, Edinburgh EH16 4SB, UK.
Roy
Douglas
D
Scottish Centre for Genomic Technology and Informatics, Medical School, University of Edinburgh, Edinburgh EH16 4SB, UK.
Forster
Thorsten
T
Scottish Centre for Genomic Technology and Informatics, Medical School, University of Edinburgh, Edinburgh EH16 4SB, UK.
Ghazal
Peter
P
Scottish Centre for Genomic Technology and Informatics, Medical School, University of Edinburgh, Edinburgh EH16 4SB, UK.
D000818:Animals; D002460:Cell Line; D020869:Gene Expression Profiling; D015967:Gene Expression Regulation, Viral; D006562:Herpes Zoster / Q000821:virology; D014645:Herpesvirus 3, Human / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis; D014158:Transcription, Genetic; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*
D014764:Viral Proteins