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American Journal of Pathology, Vol 134, 633-639, Copyright © 1989 by American Society for Investigative Pathology

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Detection of viral genomes in the liver by in situ hybridization using 35S-, bromodeoxyuridine-, and biotin-labeled probes

G Niedobitek, T Finn, H Herbst and H Stein
Department of Pathology, Klinikum Steglitz, Freie Universitat Berlin, Federal Republic of Germany.

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.




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Copyright © 1989 by the American Society for Investigative Pathology.