Serological Identification of Infectious Hematopoietic Necrosis Virus in Fixed Tissue Culture Cells by Alkaline Phosphatase Immunocytochemistry

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Abstract
An alkaline phosphatase immunocytochemical (APIC) assay was adapted for direct detection of infectious hematopoietic necrosis virus (IHNV) in infected tissue culture cells. The APIC assay provided a means of confirming the diagnosis of IHNV after the tissue culture plates had been fixed with formalin and stained with crystal violet. In cases where the original fish tissue samples had been discarded, the APIC assay was useful and was able to detect IHNV on plates that were more than 1 year old. The assay used a broadly reactive monoclonal antibody (lNDW14D) to the IHNV nucleoprotein to detect viral antigen in cells infected by virus isolates representing the five known IHNV types. Likewise, the type-2-specific monoclonal antibody (2NH105B) was able to distinguish type 2 IHNV in plaques that had been previously fixed. No cross-reactivity was seen with six other fish rhabdoviruses or with a fish birnavirus, infectious pancreatic necrosis virus (IPNV). Immunocytochernical staining was able to disting...

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