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

Journal of Aquatic Animal HealthVolume 5, Issue 4 p. 265-269 Article Serological Identification of Infectious Hematopoietic Necrosis Virus in Fixed Tissue Culture Cells by Alkaline Phosphatase Immunocytochemistry Barbara S. Drolet, Barbara S. Drolet Department of Microbiology and Laboratory for Fish Disease Research Oregon State University, Corvallis, Oregon, 97331-3804 USASearch for more papers by this authorJ. S. Rohovec, J. S. Rohovec Department of Microbiology and Laboratory for Fish Disease Research Oregon State University, Corvallis, Oregon, 97331-3804 USASearch for more papers by this authorJ. C. Leong, J. C. Leong Department of Microbiology and Laboratory for Fish Disease Research Oregon State University, Corvallis, Oregon, 97331-3804 USASearch for more papers by this author Barbara S. Drolet, Barbara S. Drolet Department of Microbiology and Laboratory for Fish Disease Research Oregon State University, Corvallis, Oregon, 97331-3804 USASearch for more papers by this authorJ. S. Rohovec, J. S. Rohovec Department of Microbiology and Laboratory for Fish Disease Research Oregon State University, Corvallis, Oregon, 97331-3804 USASearch for more papers by this authorJ. C. Leong, J. C. Leong Department of Microbiology and Laboratory for Fish Disease Research Oregon State University, Corvallis, Oregon, 97331-3804 USASearch for more papers by this author First published: 1 December 1993 https://doi.org/10.1577/1548-8667(1993)005<0265:SIOIHN>2.3.CO;2Citations: 9 To whom correspondence should be addressed. AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onEmailFacebookTwitterLinkedInRedditWechat 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 distinguish between the cytopathology produced by virus infection and that induced by the toxicity of tissue samples. In the latter case, no staining was observed. The staining was not affected by the age of the fixed and dried plates or by the polyethylene glycol pretreatment of cells. The ease, specificity, and sensitivity of the procedure made the APIC assay an attractive alternative to the standard serum neutralization or immunofluorescent identification of IHNV for diagnostic fish disease laboratories. Citing Literature Volume5, Issue41 December 1993Pages 265-269 RelatedInformation