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Pathogens. 2017 Sep 25;6(4). pii: E45. doi: 10.3390/pathogens6040045.
The Use of Hyperimmune Chicken Reference Sera Is Not Appropriate for the Validation of Influenza Pseudotype Neutralization Assays.
Ferrara F1, Molesti E2, Scott S3, Cattoli G4, Temperton N5.
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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.
KEYWORDS:
influenza; neutralization; pseudotypes; serology; standards; validation
PMID: 28946689 DOI: 10.3390/pathogens6040045
Free full text
The Use of Hyperimmune Chicken Reference Sera Is Not Appropriate for the Validation of Influenza Pseudotype Neutralization Assays.
Ferrara F1, Molesti E2, Scott S3, Cattoli G4, Temperton N5.
Author information
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.
KEYWORDS:
influenza; neutralization; pseudotypes; serology; standards; validation
PMID: 28946689 DOI: 10.3390/pathogens6040045
Free full text