Key result
In situ hybridization with biotinylated probes successfully detected human genes in male tonsil and herpes simplex virus in glial cells and neurones of herpes encephalitis brain tissue.
A simple, reproducible protocol using in situ hybridization with biotinylated probes can successfully detect human genes and viral DNA in routine formalin-fixed paraffin-embedded tissues.
Biotinylated ISH may enable gene/virus detection in routine FFPE tissues; leaves open validation before clinical adoption.
A simple reproducible protocol for detecting multiple copy human genes and viral DNA in routine formalin fixed paraffin embedded tonsil and brain, by in situ hybridisation with biotinylated probes, is described. The protocol consists of digestion of formalin fixed paraffin sections, with 0.4% pepsin in 0.01 M hydrochloric acid for one hour at 37 degrees C, followed by hybridisation with biotinylated probes. The biotinylated probes used for establishing the conditions for in situ localisation of DNA were total placental DNA (TG1), pHY 2.1 (a Y chromosome probe), and herpes simplex virus I and II. In human male tonsil TG1 labelled all nuclei and pHY 2.1 reacted only with nuclear Y bodies. In herpes encephalitis the virus was detected in some glial cells and neurones.
No takes yet. Share an insight, caveat, or question.
Burns et al. (1986) studied Herpes encephalitis. In situ hybridisation with biotinylated probes was evaluated on Detection of multiple copy human genes and viral DNA. In situ hybridization with biotinylated probes successfully detected human genes in male tonsil and herpes simplex virus in glial cells and neurones of herpes encephalitis brain tissue.
Synapse has enriched 5 closely related papers on similar clinical questions. Consider them for comparative context: