Key result
In situ hybridization successfully detected chicken anemia virus nucleic acid in thymus tissue after up to 7 days of formalin fixation, whereas immunocytochemical detection was severely impaired after 6 hours.
In situ hybridization is superior to immunocytochemistry for detecting chicken anemia virus in tissues subjected to prolonged formalin fixation.
Supports ISH for virus detection in long-fixed avian tissues; leaves open translation to other species or clinical samples.
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.
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Allan et al. (1993) studied Chicken anemia virus (CAV) infection. Biotinylated in situ hybridization vs. Immunocytochemical detection was evaluated on Detection of CAV nucleic acid/antigen after different fixation times. In situ hybridization successfully detected chicken anemia virus nucleic acid in thymus tissue after up to 7 days of formalin fixation, whereas immunocytochemical detection was severely impaired after 6 hours.
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