Two colour DNA in situ hybridization for the detection of two viral genomes using non-radioactive probes |
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Authors: | H. Mullink J. M. M. Walboomers A. K. Raap C. J. L. M. Meyer |
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Affiliation: | (1) Department of Pathology, Free University Hospital, De Boelelaan 1117, Postbus 7075, 1007 MB Amsterdam, The Netherlands;(2) Department of Cytochemistry and Cytometry, Sylvius Laboratories, University of Leiden, Wassenaarseweg 72, 2333 AL Leiden, The Netherlands |
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Abstract: | Summary 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 (HPV 16) 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 streptavidinbiotinated 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 HPV 16 DNA and haptenized HSV2 DNA. |
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