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Comparative Study
. 2016 Nov;10(6):518-524.
doi: 10.1111/irv.12408. Epub 2016 Aug 27.

A comparison of hemagglutination inhibition and neutralization assays for characterizing immunity to seasonal influenza A

Shaun Truelove  1 , Huachen Zhu  2   3 , Justin Lessler  1 , Steven Riley  4 , Jonathan M Read  5 , Shuying Wang  6 , Kin On Kwok  3 , Yi Guan  2   3 , Chao Qiang Jiang  6 , Derek A T Cummings  7   8   9
Affiliations
Comparative Study

A comparison of hemagglutination inhibition and neutralization assays for characterizing immunity to seasonal influenza A

Shaun Truelove et al. Influenza Other Respir Viruses. 2016 Nov.

Abstract

Background: Serum antibody to influenza can be used to identify past exposure and measure current immune status. The two most common methods for measuring this are the hemagglutination inhibition assay (HI) and the viral neutralization assay (NT), which have not been systematically compared for a large number of influenza viruses.

Methods: A total of 151 study participants from near Guangzhou, China, were enrolled in 2009 and provided serum. HI and NT assays were performed for 12 historic and recently circulating strains of seasonal influenza A. We compared titers using Spearman correlation and fit models to predict NT using HI results.

Results: We observed high positive mean correlation between HI and NT assays (Spearman's rank correlation, ρ=.86) across all strains. Correlation was highest within subtypes and within close proximity in time. Overall, an HI=20 corresponded to NT=10, and HI=40 corresponded to NT=20. Linear regression of log(NT) on log(HI) was statistically significant, with age modifying this relationship. Strain-specific area under a curve (AUC) indicated good accuracy (>80%) for predicting NT with HI.

Conclusions: While we found high overall correspondence of titers between NT and HI assays for seasonal influenza A, no exact equivalence between assays could be determined. This was further complicated by correspondence between titers changing with age. These findings support generalized comparison of results between assays and give further support for use of the hemagglutination inhibition assay over the more resource intensive viral neutralization assay for seasonal influenza A, although attention should be given to the effect of age on these assays.

Keywords: cross-protection; hemagglutination inhibition test; immunity; influenza; microneutralization test; neutralization test.

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Figures

Figure 1
Figure 1
Neutralization assay (NT) (left) and hemagglutination inhibition assay (HI) (right) titers for each of the 151 participants in the study plotted by rank of age (oldest at top). Color indicates the titer measured by each assay. Strains are indicated on the x‐axis of each figure
Figure 2
Figure 2
Correlation of HI and NT titers for all influenza A strains, H3N2 strains, and H1N1 strains. The orange lines indicate perfect correlation between HI and NT titers. The purple line represents the overall smoothed mean of the data
Figure 3
Figure 3
Correlations of HI and NT titers by influenza A strain. The orange lines indicate perfect correlation between HI and NT titers. The purple line represents the overall smoothed mean of the data
Figure 4
Figure 4
Sensitivity and specificity of predicting NT titer of 10, 20, and 40 using HI data for all influenza strains
See this image and copyright information in PMC

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