title: Two colour DNA in situ hybridization for the detection of two viral genomes using non-radioactive probes.
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.
authors.affiliation: Department of Pathology, Free University Hospital, Amsterdam, The Netherlands.
authors.forename: H
authors.initials: H
authors.lastname: Mullink
authors.forename: J M
authors.initials: JM
authors.lastname: Walboomers
authors.forename: A K
authors.initials: AK
authors.lastname: Raap
authors.forename: C J
authors.initials: CJ
authors.lastname: Meyer
mesh_terms: D015344:DNA Probes, HPV; D004279:DNA, Viral / Q000235:genetics*; D005260:Female; D006801:Humans; D007124:Immunoenzyme Techniques; D009693:Nucleic Acid Hybridization; D027383:Papillomaviridae / Q000235:genetics*; D018139:Simplexvirus / Q000235:genetics*; D014407:Tumor Cells, Cultured / Q000382:microbiology*; D002583:Uterine Cervical Neoplasms / Q000382:microbiology
chemical_list: D015344:DNA Probes, HPV; D004279:DNA, Viral