Hemagglutination-inhibition assay for influenza virus subtype identification and the detection and quantitation of serum antibodies to influenza virus

Methods Mol Biol. 2014:1161:11-25. doi: 10.1007/978-1-4939-0758-8_2.


Hemagglutination-inhibition (HI) assay is a classical laboratory procedure for the classification or subtyping of hemagglutinating viruses. For influenza virus, HI assay is used to identify the hemagglutinin (HA) subtype of an unknown isolate or the HA subtype specificity of antibodies to influenza virus. Since the HI assay is quantitative it is frequently applied to evaluate the antigenic relationships between different influenza virus isolates of the same subtype. The basis of the HI test is inhibition of hemagglutination with subtype-specific antibodies. The HI assay is a relatively inexpensive procedure utilizing standard laboratory equipment, is less technical than molecular tests, and is easily completed within several hours. However when working with uncharacterized viruses or antibody subtypes the library of reference reagents required for identifying antigenically distinct influenza viruses and or antibody specificities from multiple lineages of a single hemagglutinin subtype requires extensive laboratory support for the production and optimization of reagents.

MeSH terms

  • Animals
  • Antibodies, Viral / blood*
  • Antibodies, Viral / immunology*
  • Antibody Specificity
  • Antigens, Viral / immunology
  • Chickens / virology
  • Erythrocytes / virology
  • Hemagglutination Inhibition Tests / methods*
  • Hemagglutinin Glycoproteins, Influenza Virus / immunology
  • Immune Sera / blood
  • Immune Sera / immunology
  • Influenza A virus / classification*
  • Influenza A virus / immunology
  • Influenza A virus / isolation & purification*


  • Antibodies, Viral
  • Antigens, Viral
  • Hemagglutinin Glycoproteins, Influenza Virus
  • Immune Sera