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Double in situ hybridization for detection of Herpes simplex virus and cytomegalovirus DNA using non-radioactive probes

G Gentilomi, M Musiani, M Zerbini, D Gibellini, G Gallinella and S Venturoli

Institute of Microbiology, University of Bologna, Italy.

We describe a double in situ hybridization assay for the simultaneous detection of Herpes simplex virus (HSV) and cytomegalovirus (CMV) DNA in infected cell cultures using non-radioactive-labeled probes. This work used a biotinylated HSV DNA probe, which can be revealed by an avidin-biotin-peroxidase complex and a digoxigenin-labeled CMV DNA probe, visualized by anti-digoxigenin F(ab) fragments conjugated with alkaline phosphatase. Light microscopy visualization was achieved by the contrasting colors of appropriate peroxidase and alkaline phosphatase reaction products (red and dark blue, respectively). The time required to perform the double hybridization assay was about 3 hr. This double hybridization assay proved to be sensitive, specific, and provided good resolving power.

Volume 40, Issue 3, pp. 421-425, 03/01/1992
Copyright © 1992 by The Histochemical Society


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M. Musiani, M. Zerbini, S. Venturoli, G. Gentilomi, G. Gallinella, E. Manaresi, M. La Placa, A. D'Antuono, A. Roda, and P. Pasini
Sensitive Chemiluminescence In Situ Hybridization for the Detection of Human Papillomavirus Genomes in Biopsy Specimens
J. Histochem. Cytochem., May 1, 1997; 45(5): 729 - 736.
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