Two colour DNA in situ hybridization for the detection of two viral genomes using non-radioactive probes
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Updated: Oct 23, 2026

Radioactive in situ Hybridization for Detecting Diverse Gene Expression Patterns in Tissue
Published on: April 27, 2012
H Mullink1, J M Walboomers, A K Raap
1Department of Pathology, Free University Hospital, Amsterdam, The Netherlands.
Abstract:
Methods for the simultaneous detection of two virus types in cytological preparations or tissue sections by non-radioactive in situ hybridization were investigated. As a model system, CaSki cells, which have human papilloma virus type 16 (HPV16) DNA integrated in their cellular genome, were in vitro infected with Herpes simplex virus 2 (HSV2). DNA probes for both viruses were labeled with biotin, acetylaminofluorene (AAF), and transaminated-sulfonated cytosine (TS-modified). Best results were obtained when a mixture of biotinated and haptenized DNA probes (AAF- or TS-modified) was used for hybridization. The biotinated hybrid was demonstrated with a streptavidin-biotinated alkaline phosphatase staining reaction, whereas the haptenized hybrid was visualized by an indirect peroxidase method. Visualisation of both viral DNAs in the same cell was possible by a combination of biotinated HPV16 DNA and haptenized HSV2 DNA.
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