Abstract
The procedure described here was reported to detect the genomic strand of the hepatitis virus (HDV) RNA (1) in formalin-fixed, paraffin-embedded liver sections (Fig. 1). The method used a 27-mer end-labeled with digoxigenin (DIG). Hybrids were detected by a specific antibody coupled to alkaline phosphatase. Earlier reports had described the detection of both genomic and antigenomic HDV RNA by radioactive 35S-labeled RNA probes (Fig. 2) (2). However, because the average number of genomic HDV RNA in each hepatocyte is very high (around 300,000 copies/cell) (3), HDV is the ideal target to be detected by a nonradioactive in situ hybridization procedure, even using a short oligonucleotide probe, i.e., representing only approx 1.6% of the total length of the genome.
Detection of genomic HDVRNA in nuclei of hepatocytes by nonradioisotopic in situ hybridization using a digoxigenin-labeled synthetic oligonucleotide. Counterstaining with methyl green. Original magnification ×40.
Detection of genomic HDV RNA in nuclei of hepatocytes by in situ hybridization using 35S-labeled RNA probes. Counterstaining with hematoxylin. Original magnification ×100.
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Negro, F. (2004). Nonradioisotopic In Situ Hybridization for HDV RNA. In: Hamatake, R.K., Lau, J.Y.N. (eds) Hepatitis B and D Protocols. Methods in Molecular Medicine, vol 95. Humana Press. https://doi.org/10.1385/1-59259-669-X:95
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DOI: https://doi.org/10.1385/1-59259-669-X:95
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