In situ hybridisation with digoxigenin-labelled DNA probes for detection of viral genomes.
The applicability of a recently developed non-radioactive DNA labelling and detection method, which uses the digoxigenin (DIG) enzyme linked immunosorbent assay (ELISA) system, for the detection of viral infections in pathology specimens by in situ hybridisation, was examined. Its efficacy was compared with that of biotin and radioisotope labelling methods. Three cases of progressive multifocal leucoencephalopathy, two of verruca vulgaris, and seven cases of laryngeal papilloma were studied. The sensitivity of the DIG labelled probe was almost the same as that of a 35S-labelled probe in the dot-blot hybridisation test. Using in situ hybridisation with 35S-labelled and DIG labelled probes, the levels of the hybridised signals detected were similar. The biotin labelled probe was less sensitive, particularly in the cases of laryngeal papilloma. The DIG labelling and detection method was highly sensitive and applicable to the detection of viral infection by ISH, and is preferable to a radiolabelled probe, especially when in situ hybridisation is done in the pathology laboratory.
Furuta
Y
Y
Department of Pathology II, Hokkaido University School of Medicine, Sapporo, Japan.
Shinohara
T
T
Sano
K
K
Meguro
M
M
Nagashima
K
K
D001710:Biotin; D015342:DNA Probes; D003947:Diagnostic Techniques, Radioisotope; D004076:Digoxigenin; D004797:Enzyme-Linked Immunosorbent Assay; D005814:Genes, Viral; D006801:Humans; D009693:Nucleic Acid Hybridization; D027383:Papillomaviridae / Q000235:genetics; D011120:Polyomavirus / Q000235:genetics; D014412:Tumor Virus Infections / Q000175:diagnosis*
D015342:DNA Probes; D001710:Biotin; D004076:Digoxigenin