In situ amplification and detection of HIV-1 DNA in fixed pediatric AIDS brain tissue

Leroy R. Sharer*, Yoshihiro Saito, Antoinette Da Cunha, Padiwath C. Ung, Harris A. Gelbard, Leon G. Epstein, Benjamin M. Blumberg

*Corresponding author for this work

Research output: Contribution to journalArticle

34 Scopus citations

Abstract

To examine whether latent infection by HIV-1 occurs in the central nervous system, we optimized a procedure for amplification and detection of HIV-1 DNA in situ, in formalin-fixed brain tissue from a child with severe HIV-1- associated progressive encephalopathy and severe HIV-1 encephalitis. By the use of a two-step technique, which involved polymerase chain reaction with incorporation of digoxigenin-labeled nucleotides followed by in situ hybridization with biotinylated probes, we found infection of numerous mononuclear cells and astrocytes in the cerebral white matter as well as of perineuronal satellite cells in basal ganglia, but not of neurons. Following PCR amplification, nuclear signal was found in 10 to 20 times as many cells as in parallel, control experiments using conventional, unamplified in situ hybridization.

Original languageEnglish (US)
Pages (from-to)614-617
Number of pages4
JournalHuman pathology
Volume27
Issue number6
DOIs
StatePublished - Jan 1 1996

Keywords

  • AIDS encephalopathy
  • HIV-1 infection
  • in situ PCR
  • pediatric brain

ASJC Scopus subject areas

  • Pathology and Forensic Medicine

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    Sharer, L. R., Saito, Y., Da Cunha, A., Ung, P. C., Gelbard, H. A., Epstein, L. G., & Blumberg, B. M. (1996). In situ amplification and detection of HIV-1 DNA in fixed pediatric AIDS brain tissue. Human pathology, 27(6), 614-617. https://doi.org/10.1016/S0046-8177(96)90172-0