Abstract
The pseudotype particle neutralization test (pp-NT) is a next-generation serological assay employed for the sensitive study of influenza antibody responses against hemagglutinin (HA), including stalk-directed antibodies. However, a validation of this assay has yet to be performed, and this limits its use to primarily research laboratories. To identify possible serological standards to be used in optimization and validation of the pp-NT, we have evaluated the cross-reactivity of hyperimmune chicken reference antisera in this assay. Our findings show that the cross-reactivity detected by the pp-NT is only partly explained by phylogenetic relationships and protein homology between the HA subtypes analysed; further studies are necessary to understand the origin of the cross-reactivity detected, and reference standards with higher specificity should be evaluated or generated de novo for future use in pp-NT.
Highlights
Serological methods such as single radial haemolysis (SRH), haemagglutination inhibition (HI)and microneutralization (MN) are cost-effective and widely-used methodologies to monitor the circulation and prevalence of influenza viruses, and are employed in vaccine immunogenicity studies [1]
We have evaluated the neutralization activity and cross-reactivity of chicken reference antisera against a panel of pp bearing HAs of a representative strain, for each
H6 and H13 influenza pp were not used in this study since appropriate pp bearing the HA of these two subtypes were not available
Summary
Microneutralization (MN) are cost-effective and widely-used methodologies to monitor the circulation and prevalence of influenza viruses, and are employed in vaccine immunogenicity studies [1]. These assays have numerous shortcomings, especially related to the variability of the reagents, their standardization between laboratories, and their ability to detect haemagglutinin (HA) stalk-directed antibodies. The pseudotype particle neutralization test (pp-NT) appears to be more sensitive than other functional assays in the detection of the antibody response directed against the HA stalk [4]. The validation of pp-NT, which will permit its more
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