Ann Lab Med.  2012 Jul;32(4):276-282. 10.3343/alm.2012.32.4.276.

Erythrocyte Binding Preference of Human Pandemic Influenza Virus A and Its Effect on Antibody Response Detection

• Makkoch J
• Prachayangprecha S
• Payungporn S
• Chieochansin T
• Songserm T
• Amonsin A
• Poovorawan Y

• KMID: 1380087
• DOI: http://doi.org/10.3343/alm.2012.32.4.276

Abstract

BACKGROUND
Validation of hemagglutination inhibition (HI) assays is important for evaluating antibody responses to influenza virus, and selection of erythrocytes for use in these assays is important. This study aimed to determine the correlation between receptor binding specificity and effectiveness of the HI assay for detecting antibody response to pandemic influenza H1N1 (pH1N1) virus.
METHODS
Hemagglutination (HA) tests were performed using erythrocytes from 6 species. Subsequently, 8 hemagglutinating units of pH1N1 from each species were titrated by real-time reverse transcription-PCR. To investigate the effect of erythrocyte binding preference on HI antibody titers, comparisons of HI with microneutralization (MN) assays were performed.
RESULTS
Goose erythrocytes showed most specific binding with pH1N1, while HA titers using human erythrocytes were comparable to those using turkey erythrocytes. The erythrocyte binding efficiency was shown to have an impact on antibody detection. Comparing MN titers, HI titers using turkey erythrocytes yielded the most accurate results, while those using goose erythrocytes produced the highest geometric mean titer. Human blood group O erythrocytes lacking a specific antibody yielded results most comparable to those obtained using turkey erythrocytes. Further, pre-existing antibody to pH1N1 and different erythrocyte species can distort HI assay results.
CONCLUSIONS
HI assay, using turkey and human erythrocytes, yielded the most comparable and applicable results for pH1N1 than those by MN assay, and using goose erythrocytes may lead to overestimated titers. Selection of appropriate erythrocyte species for HI assay allows construction of a more reliable database, which is essential for further investigations and control of virus epidemics.

Keyword

Hemagglutination test; Hemagglutination inhibition test; Erythrocyte binding preference; Microneutralization test; H1N1 virus

MeSH Terms

Adult
Animals
Antibodies, Neutralizing/immunology
Antibodies, Viral/*analysis/immunology
Chickens
Erythrocytes/*metabolism
Female
Geese
*Hemagglutination Inhibition Tests
Horses
Humans
Influenza A Virus, H1N1 Subtype/genetics/immunology/*metabolism
Influenza, Human/epidemiology/immunology/virology
Male
Middle Aged
Neutralization Tests
Pandemics
Swine
Turkeys