In situ hybridization for the detection of chicken anemia virus in formalin-fixed, paraffin-embedded sections.
The development of a biotinylated in situ hybridization procedure for the detection of chicken anemia virus (CAV) is described. A double-stranded DNA probe was prepared using polymerase chain reaction and was biotinylated by nick translation. Hybridization conditions included the use of a microwave oven to denature target and probe nucleic acids and a five-step protocol for detection of biotinylated hybrids. In situ hybridization detected CAV nucleic acid in thymus tissue from experimentally infected birds after 6 hours, 3 days, and 7 days fixation time in formalin. However, immunocytochemical detection of CAV antigen was severely impaired if tissue was fixed in formalin for more than 6 hours.
Allan
G M
GM
Department of Agriculture, Veterinary Sciences Division, Stormont, Belfast, Northern Ireland.
Smyth
J A
JA
Todd
D
D
McNulty
M S
MS
D000740:Anemia / Q000382:microbiology / Q000662:veterinary; D000818:Animals; D001483:Base Sequence; D002645:Chickens / Q000382:microbiology*; D004267:DNA Viruses / Q000302:isolation & purification*; D005557:Formaldehyde; D007150:Immunohistochemistry; D017403:In Situ Hybridization / Q000379:methods / Q000662:veterinary*; D008969:Molecular Sequence Data; D016612:Paraffin Embedding / Q000662:veterinary; D011201:Poultry Diseases / Q000382:microbiology; D013950:Thymus Gland / Q000382:microbiology; D016707:Tissue Fixation / Q000662:veterinary; D014777:Virus Diseases / Q000662:veterinary
D005557:Formaldehyde