title: A hybrido-immunocytochemical assay for the in situ detection of cytomegalovirus DNA using digoxigenin-labeled probes.
abstract: A non-radioactive hybrido-immunocytochemical assay for the detection of cytomegalovirus (CMV) DNA in infected cells was developed. Two different DNA fragments belonging to the repeated sequences of CMV genome were used to construct the hybridization probe. The probe was constructed by incorporating deoxyuridine triphosphate labeled with digoxigenin. The in situ hybridized CMV DNA probe was immunocytochemically visualized by anti-digoxigenin. Fab fragments labeled with alkaline phosphatase. This procedure permitted the DNA detection, in the nuclei of infected cells fixed at 48 h after infection, of the Towne CMV reference strain and 21 different laboratory-isolated CMV strains. Our assay demonstrated a high specificity, sensitivity and reproducibility.
authors.affiliation: Institute of Microbiology, University of Bologna, Italy.
authors.forename: G
authors.initials: G
authors.lastname: Gentilomi
authors.forename: M
authors.initials: M
authors.lastname: Musiani
authors.forename: M
authors.initials: M
authors.lastname: Zerbini
authors.forename: G
authors.initials: G
authors.lastname: Gallinella
authors.forename: D
authors.initials: D
authors.lastname: Gibellini
authors.forename: M
authors.initials: M
authors.lastname: La Placa
mesh_terms: D002478:Cells, Cultured; D003587:Cytomegalovirus / Q000032:analysis*; D003586:Cytomegalovirus Infections / Q000175:diagnosis*; D015342:DNA Probes; D004279:DNA, Viral / Q000032:analysis*; D004076:Digoxigenin; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D009693:Nucleic Acid Hybridization
chemical_list: D015342:DNA Probes; D004279:DNA, Viral; D004076:Digoxigenin