A non-destructive test for detection of IPNV-carriers in Atlantic halibut, Hippoglossus hippoglossus (L.).
Over 18 months after infectious pancreatic necrosis virus (IPNV) was first detected in fish (80 g-4 kg) on a halibut farm, the stocks were found to be still carrying the virus. This suggests that long-term persistence of IPNV occurs in farmed Atlantic halibut. A non-destructive test was applied to blood adherent leucocytes by placing 100 microL of whole blood collected in a heparinized tube into 96-well plates. After overnight incubation, the non-adherent cells were washed off, the remaining adherent cells lysed in a lysis buffer and inoculated onto CHSE-214 cells. The resulting cytopathic effect was confirmed as IPNV positive by enzyme-linked immunosorbent assay. In a sample of 10 fish tested by this method, all were positive for IPNV while only two were positive by the standard method for virus culture from sonicated kidney homogenates and only one fish, which was positive by the standard method, was positive by reverse transcription polymerase chain reaction on kidney tissue. The test on blood leucocytes is shown to be simple to perform on samples taken under field conditions.
Gahlawat
S K
SK
Department of Zoology and Aquaculture, CCS Haryana Agricultural University, Hisar, India.
Munro
E S
ES
Ellis
A E
AE
D000818:Animals; D017756:Aquaculture / Q000379:methods; D018175:Birnaviridae Infections / Q000175:diagnosis / Q000662:veterinary*; D002478:Cells, Cultured; D017931:DNA Primers; D004797:Enzyme-Linked Immunosorbent Assay; D005393:Fish Diseases / Q000175:diagnosis* / Q000821:virology*; D005399:Fishes; D018061:Infectious pancreatic necrosis virus / Q000302:isolation & purification*; D007668:Kidney / Q000821:virology; D007962:Leukocytes / Q000821:virology; D008264:Macrophages / Q000821:virology; D011205:Povidone; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012822:Silicon Dioxide; D010948:Viral Plaque Assay / Q000379:methods*
D017931:DNA Primers; C016039:Percoll; D012822:Silicon Dioxide; D011205:Povidone