Detection of bovine virus diarrhoea virus RNA by in situ hybridisation with digoxigenin-labelled riboprobes.
A non-isotopic in situ hybridisation (ISH) method has been developed for the detection of bovine virus diarrhoea virus (BVDV) RNA. The technique is highly specific and allows the detection of BVDV in formalin-fixed paraffin-wax-embedded sections. Digoxigenin-labelled riboprobes were generated to the coding region for the p125 non-structural protein and consequently it was possible to locate the virus positive-sense RNA in BVDV-infected tissues. Formalin-fixed tissues are ideal for ISH, combining the possibility of a retrospective analysis of tissues for virus distribution with excellent morphological preservation. In situ hybridisation has facilitated a precise examination, at the cellular level, of viral nucleic acid distribution during pestiviral infections, with an increased sensitivity and wider applicability than immunocytochemistry.
Desport
M
M
Institute for Animal Health, Compton, Newbury, Berks, UK.
Collins
M E
ME
Brownlie
J
J
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000956:Antigens, Viral / Q000032:analysis; D001912:Bovine Virus Diarrhea-Mucosal Disease / Q000821:virology*; D002417:Cattle; D002540:Cerebral Cortex / Q000276:immunology / Q000821:virology; D001908:Diarrhea Viruses, Bovine Viral / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D004076:Digoxigenin; D019286:Endopeptidase K; D005260:Female; D007150:Immunohistochemistry; D017403:In Situ Hybridization / Q000379:methods*; D008198:Lymph Nodes / Q000276:immunology / Q000821:virology; D014066:Palatine Tonsil / Q000276:immunology / Q000821:virology; D015347:RNA Probes; D012367:RNA, Viral / Q000032:analysis*; D012697:Serine Endopeptidases
D000911:Antibodies, Monoclonal; D000956:Antigens, Viral; D015347:RNA Probes; D012367:RNA, Viral; D012697:Serine Endopeptidases; D019286:Endopeptidase K; D004076:Digoxigenin