In situ hybridization: an optimised detection protocol for a biotinylated DNA probe renders it more sensitive than a comparable 35S-labelled probe.
The development of a highly sensitive biotinylated in situ hybridization protocol using egg drop syndrome virus infection of domestic fowl as a model is described. This protocol incorporates the use of a monoclonal antibody to biotin as the initial step in a peroxidase-based detection system. The detection of viral nucleic acid in formalin-fixed, paraffin-embedded sections using biotinylated and 35S-labelled probes is compared to the detection of viral antigen by immunocytochemistry. Biotinylated probes detected more positive cells than 35S-labelled probes and were more specific. Biotinylated probes detected more positive cells than immunocytochemistry.
Allan
G M
GM
Veterinary Research Laboratories, Stormont, Belfast, Ireland, U.K.
Todd
D
D
Smyth
J A
JA
Mackie
D P
DP
Burns
J
J
McNulty
M S
MS
D000256:Adenoviridae / Q000302:isolation & purification*; D000257:Adenoviridae Infections / Q000175:diagnosis*; D000818:Animals; D000956:Antigens, Viral / Q000032:analysis*; D000259:Aviadenovirus / Q000235:genetics / Q000302:isolation & purification*; D001710:Biotin; D002645:Chickens; D015342:DNA Probes; D004279:DNA, Viral / Q000032:analysis*; D004195:Disease Models, Animal; D007124:Immunoenzyme Techniques; D011933:Reagent Kits, Diagnostic; D012680:Sensitivity and Specificity; D013462:Sulfur Radioisotopes
D000956:Antigens, Viral; D015342:DNA Probes; D004279:DNA, Viral; D011933:Reagent Kits, Diagnostic; D013462:Sulfur Radioisotopes; D001710:Biotin