Comparison of hemagglutination inhibition, single radial hemolysis, virus neutralization assays, and ELISA to detect antibody levels against seasonal influenza viruses

Influenza Other Respir Viruses. 2018 Nov;12(6):675-686. doi: 10.1111/irv.12591. Epub 2018 Aug 11.

Abstract

Background: The immunological response to influenza vaccine and/or natural infection is evaluated by serological techniques, the most common being hemagglutination inhibition (HI), single radial hemolysis (SRH), and virus neutralization assays, which is commonly used in a micro-neutralization (MN) format. ELISA is not officially required; however, this assay is able to measure different class-specific antibodies. The four assays identify different sets or subsets of antibodies.

Objectives: The aim of this study was to establish the correlation among four serological assays using four seasonal influenza strains.

Methods: The HI, SRH, MN assays, and ELISA were performed on four seasonal influenza strains.

Results: A strong positive correlation was found between HI and MN and between SRH and MN assays for influenza A strains. The B strains also showed good correlations among the three assays. A positive correlation was also found between ELISA and the "classical" assays for all strains. Concerning the correlates of protection, as defined by HI ≥ 40 and SRH ≥ 25 mm2 , good agreement was observed for the influenza A strains. By contrast, the agreement for the B strains was very low.

Conclusions: There is a positive strong correlation among the four serological assays for both A and B strains, especially for the HI and MN assays. There is good agreement on correlates of protection between HI and SRH assays for the A strains, but very low agreement for the B strains, suggesting higher sensitivity of SRH than HI assay in detecting antibodies against the influenza B viruses.

Keywords: ELISA; correlates of protection; hemagglutination inhibition assay; single radial hemolysis; virus neutralization assay.

Publication types

  • Comparative Study
  • Evaluation Study

MeSH terms

  • Adult
  • Antibodies, Neutralizing / blood*
  • Antibodies, Viral / blood*
  • Humans
  • Immunoassay / methods*
  • Influenza, Human / immunology*
  • Orthomyxoviridae / immunology*
  • Sensitivity and Specificity

Substances

  • Antibodies, Neutralizing
  • Antibodies, Viral