Detection of viral genomes in the liver by in situ hybridization using 35S-, bromodeoxyuridine-, and biotin-labeled probes.
Methods employing 35S-, biotin-, and bromodeoxyuridine (BrdUrd)-labeled DNA probes were compared for the detection of hepatitis B virus (HBV) and cytomegalovirus (CMV) in the liver. The results demonstrate that: 1) HBV can be detected reliably only by the use of radiolabeled probes, whereas methods employing nonradioactive probes obviously are not sensitive enough for this virus. The use of 35S-labeled probes shortens the exposure times considerably in comparison to tritiated probes. 2) Biotin-labeled probes are of limited value for in situ hybridization on liver tissues because the presence of endogenous avidin-binding activity often leads to false positive results. 3) Brd-Urd-labeled probes are a useful alternative to biotinylated probes for the detection of CMV. In comparison with biotinylated probes, BrdUrd-labeled probes produce a specific signal of similar staining intensity in the absence of background staining in the liver.
Niedobitek
G
G
Department of Pathology, Klinikum Steglitz, Freie Universität Berlin, Federal Republic of Germany.
Finn
T
T
Herbst
H
H
Stein
H
H
D001710:Biotin; D001973:Bromodeoxyuridine; D003587:Cytomegalovirus / Q000235:genetics; D015342:DNA Probes; D004279:DNA, Viral; D005814:Genes, Viral; D006512:Hepatitis B Core Antigens / Q000032:analysis; D006514:Hepatitis B Surface Antigens / Q000032:analysis; D006515:Hepatitis B virus / Q000235:genetics; D006801:Humans; D007150:Immunohistochemistry; D008099:Liver / Q000276:immunology / Q000502:physiology*; D009693:Nucleic Acid Hybridization; D013462:Sulfur Radioisotopes
D015342:DNA Probes; D004279:DNA, Viral; D006512:Hepatitis B Core Antigens; D006514:Hepatitis B Surface Antigens; D013462:Sulfur Radioisotopes; D001710:Biotin; D001973:Bromodeoxyuridine