Detection of highly pathogenic influenza and pandemic influenza virus in formalin fixed tissues by immunohistochemical methods

J Virol Methods. 2012 Feb;179(2):409-13. doi: 10.1016/j.jviromet.2011.11.006. Epub 2011 Nov 18.

Abstract

Tissues infected with highly pathogenic avian influenza viruses such as H5N1 and H7N7 are normally required to be fixed in formalin or paraformaldehyde before examination in order to inactivate the virus. In this study commercially available monoclonal antibodies to the influenza nucleoprotein (NP) were evaluated in order to determine which antibodies would identify positive cells in tissues fixed in formalin or paraformaldehyde. An assessment of which antigen retrieval process would unmask antigens blocked by formalin fixation was also made. Of six commercially available monoclonal antibodies tested, only one (HB65, European Veterinary Laboratories) was able to identify all formalin fixed avian, swine and human influenza virus infected tissues, and this was after pronase induced epitope retrieval. This monoclonal antibody is recommended for routine diagnostic use for the detection of influenza A infected tissues that have been fixed in formalin or paraformaldehyde.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal
  • Antibodies, Viral
  • Birds
  • Fixatives / pharmacology
  • Formaldehyde / pharmacology
  • Humans
  • Immunohistochemistry / methods*
  • Influenza in Birds / virology
  • Influenza, Human / virology
  • Nucleocapsid Proteins
  • Orthomyxoviridae / isolation & purification*
  • Paramyxoviridae Infections / veterinary
  • Paramyxoviridae Infections / virology
  • Pathology / methods*
  • Polymers / pharmacology
  • RNA-Binding Proteins / analysis
  • RNA-Binding Proteins / immunology
  • Swine
  • Swine Diseases / virology
  • Tissue Fixation / methods
  • Viral Core Proteins / analysis
  • Viral Core Proteins / immunology

Substances

  • Antibodies, Monoclonal
  • Antibodies, Viral
  • Fixatives
  • NP protein, Influenza A virus
  • Nucleocapsid Proteins
  • Polymers
  • RNA-Binding Proteins
  • Viral Core Proteins
  • Formaldehyde
  • paraform