Optimization of an in situ hybridization technique for the detection of foot-and-mouth disease virus in bovine tissues using the digoxigenin system.
An in situ hybridization technique has been optimised for use on paraffin-embedded sections of tissues collected from cattle infected experimentally with foot-and-mouth disease virus type O1BFS. Tissue was collected 5 days after infection by direct contact. In situ hybridization was carried out using an RNA probe corresponding to a region of the 3D gene which codes for the RNA polymerase, and labelled with digoxigenin. Consistent, reproducible signal was detected within the epithelial layers of the palatine tonsil, soft palate and pharyngeal tissue studied. This is the first time that a digoxigenin-based system has been used successfully for FMD virus RNA detection with bovine tissue samples.
Woodbury
E L
EL
Institute for Animal Health, Pirbright Laboratory, Woking, Surrey, UK.
Ilott
M C
MC
Brown
C C
CC
Salt
J S
JS
D000818:Animals; D005537:Aphthovirus / Q000201:enzymology / Q000235:genetics* / Q000302:isolation & purification*; D002417:Cattle; D002418:Cattle Diseases / Q000821:virology; D004076:Digoxigenin; D005069:Evaluation Studies as Topic; D005536:Foot-and-Mouth Disease / Q000821:virology; D005814:Genes, Viral; D017403:In Situ Hybridization / Q000379:methods / Q000662:veterinary*; D010160:Palate, Soft / Q000821:virology; D014066:Palatine Tonsil / Q000821:virology; D010614:Pharynx / Q000821:virology; D015347:RNA Probes; D012324:RNA-Dependent RNA Polymerase / Q000235:genetics; D014773:Virology / Q000379:methods
D015347:RNA Probes; D012324:RNA-Dependent RNA Polymerase; D004076:Digoxigenin