Polyacrylamide gel electrophoresis of viral genomic RNA as a diagnostic method for infectious pancreatic necrosis virus detection.
A rapid and simple technique for the diagnosis of IPNV from cell culture and infected fish tissues has been developed. It is based on the observation of IPNV bisegmented double-stranded RNA genome in silver stained polyacrylamide gels after electrophoresis. The method is highly specific and can detect as little as 1 ng of viral RNA, which corresponds to 1 x 10(5) pfu, making it possible to visualize the viral genome as soon as the initial cythopatic effect appears. Furthermore, the RNA viral genome could be detected directly from fish tissues when fish showed clear clinical signs of the disease. The method has the advantage of detecting any strain of IPNV. An acrylamide concentration of 6% and bisacrylamide concentration of 0.13% give a rapid and definitive result in less than 6 h.
Ganga
M A
MA
Unidad de Virología, Instituto de Nutrición Y Tecnología de los Alimentos (INTA), Universidad de Chile, Santiago 11.
González
M P
MP
López-Lastra
M
M
Sandino
A M
AM
D000818:Animals; D018175:Birnaviridae Infections / Q000662:veterinary*; D002478:Cells, Cultured; D004591:Electrophoresis, Polyacrylamide Gel; D005393:Fish Diseases / Q000821:virology*; D005399:Fishes; D016679:Genome, Viral; D018061:Infectious pancreatic necrosis virus / Q000235:genetics / Q000302:isolation & purification*; D012367:RNA, Viral / Q000302:isolation & purification*
D012367:RNA, Viral