High-Sensitivity In Situ Hybridization DNAscope for Detection of Papillomavirus or Epstein-Barr Virus Genomes Combined with Immunofluorescent Staining in Adherent Cells.

Hybridization methods have been broadly used for the observation of DNA of different origins. Herein, we describe the highly effective and specific in situ DNA hybridization protocol DNAscope for the identification of viral genomes in cells growing in monolayer cultures. The procedure utilizes the Z-probes and hybridization-based fluorescent detection systems available from Advanced Cell Diagnostics, ACD (Newark, CA, USA). We developed our protocol for the identification of episomal and integrated human papillomavirus (HPV) or Epstein-Barr virus (EBV) DNA in human keratinocytes. This procedure can be easily adapted to the detection of the wide range of DNA viruses in cell cultures or patient-derived samples. The protocol described here combines the detection of viral genomes by DNAscope with the visualization of the cellular protein expression with immunofluorescence staining (IF).