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

Claudia Maria Trombetta, Edmond J Remarque, Daniella Mortier, Emanuele Montomoli, Claudia Maria Trombetta, Edmond J Remarque, Daniella Mortier, Emanuele Montomoli

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.

© 2018 The Authors. Influenza and Other Respiratory Viruses Published by John Wiley & Sons Ltd.

Figures

Figure 1
Figure 1
Correlation between HI‐MN, HI‐SRH, and SRH‐MN titers. Size and color indicate the number of observations at the same coordinates. The size of the circle is proportional to the square root of the number of observations at that position; thus, the size is directly proportional to the number of observations. Symbol colors also indicate the number of observations (magma color), where black indicates single observations and yellow indicates many observations. A, . B, . C,
Figure 2
Figure 2
Correlation between ELISA (IgG titer)‐HI, ELISA (IgG titer)‐MN, and ELISA (IgG titer)‐SRH titers. The points are plotted in such a way as to show where the majority of observations are located

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