Detection of infectious laryngotracheitis virus in chickens using a non-radioactive DNA probe.
A DNA hybridization assay using a non-radioactive probe has been developed for the detection of infectious laryngotracheitis virus (ILTV) DNA. A 1.4-kilobase pair BamHI fragment of ILTV genomic DNA was cloned and then labeled by one of two methods; nick translation using 32P-dATP or non-radioactive labeling using a commercially available DNA labeling and detection kit. The non-radioactive DNA labeling method proved to be as sensitive as the radioactive method. Using the non-radioactive probe, ILTV DNA was readily detected in tracheal samples from acutely infected chickens and also from convalescent chickens at a time when viral antigen could no longer be detected by the enzyme-linked immunosorbent assay or the virus could no longer be reisolated. This technique provides a safe and effective means of identifying field outbreaks of ILTV and also may detect latent ILTV infections relatively quickly and inexpensively.
Keam
L
L
Commonwealth Scientific and Industrial Research Organisation, Division of Animal Health, Parkville, Victoria, Australia.
York
J J
JJ
Sheppard
M
M
Fahey
K J
KJ
D000818:Animals; D000956:Antigens, Viral / Q000032:analysis; D002645:Chickens; D003001:Cloning, Molecular; D015342:DNA Probes; D004279:DNA, Viral / Q000032:analysis*; D004797:Enzyme-Linked Immunosorbent Assay; D006566:Herpesviridae Infections / Q000175:diagnosis / Q000382:microbiology / Q000662:veterinary*; D001352:Herpesvirus 1, Gallid / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D009693:Nucleic Acid Hybridization; D011201:Poultry Diseases / Q000175:diagnosis / Q000382:microbiology*; D011237:Predictive Value of Tests; D013047:Specific Pathogen-Free Organisms; D014132:Trachea / Q000382:microbiology
D000956:Antigens, Viral; D015342:DNA Probes; D004279:DNA, Viral