Dot blot hybridization assay for chicken anemia agent using a cloned DNA probe.
A dot blot hybridization assay capable of detecting chicken anemia agent (CAA)-specific DNA in tissues from infected birds has been developed. The assay uses a 32P-labeled DNA probe prepared from cloned CAA-specific fragments representing the entire virus genome and has a sensitivity limit of between and 1 and 10 pg. DNAs from CAA isolates originating in the Federal Republic of Germany, Japan, the United States, the United Kingdom, and Australia were detected. Investigation of specimens from experimentally infected chicks indicated that virus-specific DNA was detected in the tissues of birds from 5 through 42 days after infection and that greater amounts were usually detected in the thymus than in the spleen, liver, feces, or blood. Tissues from specific-pathogen-free and broiler chicks which had become infected at an older age through contact with experimentally infected anemic chicks also contained CAA-specific DNA detectable by the assay. Thymuses from 1- to 2-week-old chicks from eight commercial broiler flocks which had been showing clinical signs characteristic of anemia-dermatitis syndrome were found positive by the hybridization technique, but thymuses from chicks obtained from broiler flocks which did not show such signs were found negative. Of the 35 positive samples (from 46 samples tested), 19 (54%) contained virus-specific DNA in sufficiently great amounts to permit 4-h autoradiography exposures and sample throughput times of 2 days. When compared with virus isolation, the CAA dot blot hybridization assay is time- and labor-saving.
Todd
D
D
Veterinary Research Laboratories, Stormont, Belfast, United Kingdom.
Creelan
J L
JL
McNulty
M S
MS
D000740:Anemia / Q000382:microbiology*; D000818:Animals; D002645:Chickens; D015342:DNA Probes; D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification; D005069:Evaluation Studies as Topic; D015151:Immunoblotting / Q000379:methods*; D009693:Nucleic Acid Hybridization; D012680:Sensitivity and Specificity; D014773:Virology / Q000379:methods; D014780:Viruses / Q000235:genetics* / Q000302:isolation & purification
D015342:DNA Probes; D004279:DNA, Viral