Amplification and detection of lentiviral DNA inside cells

Ashley T. Haase, Ernest F. Retzel, Katherine A. Staskus

Research output: Contribution to journalArticlepeer-review

230 Scopus citations

Abstract

Visna virus and human immunodeficiency virus are prototypes of animal and human lentiviruses, respectively, that persist and are disseminated despite the host immune response because cells in the tissues and the blood-stream harbor viral genomes in a covert state. To facilitate identification of these latently infected cells, the polymerase chain reaction has been adapted to amplify viral DNA in fixed cells for detection by in situ hybridization. By using a multiple primer set that generates DNA segments with overlapping cohesive termini, visna virus DNA can be amplified, retained, and detected in infected cells with sensitivities that exceed those of existing methods by more than 2 orders of magnitude. This advance in single-cell technology should prove useful in diagnosing and gaining insight into the pathogenesis of viral infections and provide new opportunities to look for viruses in chronic diseases of unknown etiology.

Original languageEnglish (US)
Pages (from-to)4971-4975
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume87
Issue number13
DOIs
StatePublished - 1990

Keywords

  • In situ hybridization
  • Polymerase chain reaction
  • Visna virus

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