Detection of infectious pancreatic necrosis virus (IPNV) RNA by hybridization with an oligonucleotide DNA probe.
Marine farming of Atlantic salmon (Salmo salar) is growing rapidly in Norway, and fish diseases have now become one of the biggest obstacles for this new industry. Infectious pancreatic necrosis virus (IPNV) is commonly found in fish from Norwegian sea farms. Diagnosis of IPNV infection is, at present, mainly based on virus isolation in cell cultures and identification by neutralization tests (NT). A DNA-RNA hybridization assay was developed using a 24 base DNA oligonucleotide probe. This is homologous to a part of the nucleotide sequence of the IPNV genome coding for a protease. RNA extracted from IPNV and harvest from infected cell cultures were fixed to nylon filters and hybridized with the 32P end-labelled probe. The results showed that the probe specifically identified IPNV from these two materials, both for the three different virus strains (Ab, Sp and VR-299) used, and for several different field isolates. It did not hybridize with reoviruses or non-infected cell cultures used as controls. These results indicate that the probe is not serotype specific, and furthermore that RNA extraction is not required before hybridization. This method may be a useful alternative to NT for routine identification of IPNV, particularly when non-radioactive labelling of the probe is introduced.
Rimstad
E
E
Department of Microbiology and Immunology, Norwegian College of Veterinary Medicine, Oslo.
Krona
R
R
Hornes
E
E
Olsvik
O
O
Hyllseth
B
B
D000818:Animals; D001345:Autoradiography; D015342:DNA Probes; D005393:Fish Diseases / Q000175:diagnosis / Q000382:microbiology*; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes; D012367:RNA, Viral / Q000032:analysis*; D012087:Reoviridae / Q000235:genetics / Q000302:isolation & purification*; D012088:Reoviridae Infections / Q000175:diagnosis / Q000382:microbiology / Q000662:veterinary*; D012474:Salmon
D015342:DNA Probes; D015345:Oligonucleotide Probes; D012367:RNA, Viral