The specific detection of infectious pancreatic necrosis virus in infected cells and fish by the immuno dot blot method

Abstract

Abstract. The immuno dot blot technique is a rapid, sensitive and quantitative method for the diagnosis of infectious pancreatic necrosis virus (ipnv) in both infected tissue culture lysates and tissue homogenates of fish. Uninfected tissue culture lysates and fish homogenates do not react in the peroxidase dot blot system after appropriate treatment. The immuno dot blot method takes 4h to complete, and the dot blot membrane can be stored in the dark for a long period. The viral proteins in the infected tissue culture cells could be detected at 3h post‐infection if an initial high MOI was used. The minimum dose of ipnv‐ab strain which could be detected using homologous serum was 2·5 ng purified ipnv protein/dot. The smallest infectious dose of VR‐299, SP, AB, EVE and CY‐1 ipnv which could be detected by the immuno dot blot system was 104 to 105 TCID50. When one μg of purified ipnv was used, a 1·6 × 104 dilution of rabbit anti‐AB antisera could be detected.