title: Optimization of an enzyme-linked lectin assay suitable for rapid antigenic characterization of the neuraminidase of human influenza A(H3N2) viruses.
abstract: Antibodies to neuraminidase (NA), the second most abundant surface protein of the influenza virus, contribute to protection against influenza virus infection. Although traditional and miniaturized thiobarbituric acid (TBA) neuraminidase inhibition (NI) assays have been successfully used to characterize the antigenic properties of NA, these methods are cumbersome and not easily amendable to rapid screening. An additional difficulty of the NI assay is the interference by hemagglutinin (HA)-specific antibodies. To prevent interference of HA-specific antibodies, most NI assays are performed with recombinant viruses containing a mismatched HA. However, generation of these viruses is time consuming and unsuitable for large-scale surveillance. The feasibility of using the recently developed enzyme-linked lectin assay (ELLA) to evaluate the antigenic relatedness of NA of wild type A(H3N2) viruses was assessed. Rather than using recombinant viruses, wild type A(H3N2) viruses were used as antigen with ferret sera elicited against recombinant viruses with a mismatched HA. In this study, details of the critical steps that are needed to modify and optimize the NI ELLA in a format that is reproducible, highly sensitive, and useful for influenza virus surveillance to monitor antigenic drift of NA are provided.
authors.affiliation: Department of Viroscience, Erasmus Medical Center, Rotterdam, The Netherlands.
authors.forename: Kim B
authors.initials: KB
authors.lastname: Westgeest
authors.affiliation: Department of Viroscience, Erasmus Medical Center, Rotterdam, The Netherlands.
authors.forename: Theo M
authors.initials: TM
authors.lastname: Bestebroer
authors.affiliation: Department of Viroscience, Erasmus Medical Center, Rotterdam, The Netherlands.
authors.forename: Monique I J
authors.initials: MI
authors.lastname: Spronken
authors.affiliation: Division of Viral Products, Center for Biologics Evaluation and Research, Food and Drug Administration, Silver Spring, MD 20993, USA.
authors.forename: Jin
authors.initials: J
authors.lastname: Gao
authors.affiliation: Division of Viral Products, Center for Biologics Evaluation and Research, Food and Drug Administration, Silver Spring, MD 20993, USA.
authors.forename: Laura
authors.initials: L
authors.lastname: Couzens
authors.affiliation: Department of Viroscience, Erasmus Medical Center, Rotterdam, The Netherlands.
authors.forename: Albert D M E
authors.initials: AD
authors.lastname: Osterhaus
authors.affiliation: Division of Viral Products, Center for Biologics Evaluation and Research, Food and Drug Administration, Silver Spring, MD 20993, USA.
authors.forename: Maryna
authors.initials: M
authors.lastname: Eichelberger
authors.affiliation: Department of Viroscience, Erasmus Medical Center, Rotterdam, The Netherlands.
authors.forename: Ron A M
authors.initials: RA
authors.lastname: Fouchier
authors.affiliation: Department of Viroscience, Erasmus Medical Center, Rotterdam, The Netherlands. Electronic address: m.degraaf@erasmusmc.nl.
authors.forename: Miranda
authors.initials: M
authors.lastname: de Graaf
mesh_terms: D006801:Humans; D053122:Influenza A Virus, H3N2 Subtype / Q000737:chemistry* / Q000145:classification* / Q000201:enzymology; D037102:Lectins / Q000378:metabolism*; D009439:Neuraminidase / Q000032:analysis* / Q000145:classification*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013997:Time Factors; D014764:Viral Proteins / Q000032:analysis* / Q000145:classification*; D014773:Virology / Q000379:methods*
chemical_list: D037102:Lectins; D014764:Viral Proteins; C487630:NA protein, influenza A virus; D009439:Neuraminidase
keywords: Antigenic drift; ELLA; Influenza; Neuraminidase