Analysis of persistent virus infections by in situ hybridization to whole-mouse sections

Peter J. Southern, Paul Blount, Michael B.A. Oldstone

Research output: Contribution to journalArticle

41 Scopus citations

Abstract

Nucleic acid hybridization techniques have contributed significantly to the understanding of gene organization, regulation and expression1-4. In the context of persistent or latent viral infections, hybridization with specific labelled probes represents the most sensitive assay presently available for detection of viral genomes5-8. During the course of persistence, viral genomes may exist in multiple and yet quite segregated areas in an infected host, but examination of all tissues remains difficult and time-consuming. At present, the application of hybridization assays to in vivo infections requires either chemical extraction of nucleic acid coupled with dot-blot9 and gel transfer1 techniques or in situ hybridization5,10 to cryostat tissue sections. In both cases, selected tissues must be removed by dissection before analysis. We describe here a procedure which allows efficient and reproducible screening of all tissue in an infected host. Our technique allows detection of viral genetic material in whole-body sections of infected mice, and provides the first evidence in vivo for accumulation of viral genetic material with a parallel decrease in infectious virus during persistent virus infection. This technique should be widely applicable to studies of developmental regulation of gene expression, for monitoring locations of gene expression in transgenic mice and for analysis of molecular mechanisms in pathogenesis.

Original languageEnglish (US)
Pages (from-to)555-558
Number of pages4
JournalNature
Volume312
Issue number5994
DOIs
StatePublished - Dec 1 1984

ASJC Scopus subject areas

  • General

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