Abstract
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.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Beard CW (1980) Hemagglutination-inhibition. In: Hitcher SB, Domermuth CH, Purchase HG, Williams JE (eds) Isolation and identification of avian pathogens. American Association of Avian Pathologists, Kennett Square, PA, pp 331–336
Centers for Disease Control and Prevention (1982) Concepts and procedures for laboratory based influenza surveillance. CDC, U.S. Department of Health and Human Services, Washington, DC
Palmer DF, Coleman MT, Dowdle WD, Schild GO (1975) Advanced laboratory techniques for influenza diagnosis. In: Immunology series no. 6. U.S. Department of Health, Education and Welfare, Public Health Service, CDC, Atlanta, GA
Swayne DE, Halvorson DA (2003) Influenza. In: Saif YM, Barnes HJ, Gilsson JR, Fadly AM, McDougald LR, Swayne DE (eds) Diseases of poultry, 11th edn. Iowa State University Press, Ames, IA, pp 135–155
Kendal AP (1982) New techniques in antigenic analysis with influenza viruses. In: Beare AS (ed) Basic and applied influenza research. CRC, Boca Raton, FL, pp 51–78
Lee CW, Senne DA, Suarez DL (2006) Development and application of reference antisera against 15 hemagglutinin subtypes of influenza by DNA vaccination of chickens. Clin Vaccine Immunol 13:395–402
Easterday BC, Hinshaw VS, Halvorson DA (1977) Influenza. In: Calnek BW, Barnes HJ, Beard CW, McDougald LR, Saif YM (eds) Diseases of poultry, 10th edn. Iowa State University Press, Ames, IA, pp 583–605
Spackman E, Senne DA, Myers TJ, Bulaga LL, Garber LP, Perdue ML, Lohman K, Daum LT, Suarez DL (2002) Development of a real-time reverse transcriptase PCR assay for type A influenza virus and the avian H5 and H7 hemagglutinin subtypes. J Clin Microbiol 40:3256–3260
Brugh M, Beard CW (1980) Collection and processing of blood samples dried on paper for microassay of Newcastle disease virus and avian influenza virus antibodies. Am J Vet Res 41:1495–1498
Barrett T, Inglis SC (1985) Growth, purification and titration of influenza viruses. In: Mahy BWJ, Rickwood D, Hames BD (eds) Virology, a practical approach. IRL, Oxford, pp 119–150
Hanson RP, Appleton GS, Chute HL, Grumbles LC, Hitchner SB, Peacock GV, Pomeroy BS, Rosenwald AS (1971) Identification and differentiation of avian viruses. In: Methods for examining poultry biologics and for identifying and quantifying avian pathogens. National Academy of Sciences, p 261–290
Mahy BWJ, Kangro H (1996) Hemagglutination-inhibition. In: Virology methods manual, Academic, London, p 114–116
World Animal Health Organization (OIE) (2012) AvianInfluenza. In: Manual of standards for diagnostic tests and vaccines. http://www.oie.int/fileadmin/Home/eng/Health_standards/tahm/2.03.04_AI.pdf. Accessed 1 June 2013
Swayne DE, Senne DA, Beard CW (1998) Avian influenza. In: Dufour-Zavala L (ed) A laboratory manual for the isolation and identification of avian pathogens, 4th edn. American Association of Avian Pathologists, Kennett Square, PA, pp 128–134
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2014 Springer Science+Business Media New York
About this protocol
Cite this protocol
Pedersen, J.C. (2014). Hemagglutination-Inhibition Assay for Influenza Virus Subtype Identification and the Detection and Quantitation of Serum Antibodies to Influenza Virus. In: Spackman, E. (eds) Animal Influenza Virus. Methods in Molecular Biology, vol 1161. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0758-8_2
Download citation
DOI: https://doi.org/10.1007/978-1-4939-0758-8_2
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-0757-1
Online ISBN: 978-1-4939-0758-8
eBook Packages: Springer Protocols