title: Detection of influenza antigenic variants directly from clinical samples using polyclonal antibody based proximity ligation assays.
abstract: Identification of antigenic variants is the key to a successful influenza vaccination program. The empirical serological methods to determine influenza antigenic properties require viral propagation. Here a novel quantitative PCR-based antigenic characterization method using polyclonal antibody and proximity ligation assays, or so-called polyPLA, was developed and validated. This method can detect a viral titer that is less than 1000 TCID50/mL. Not only can this method differentiate between different HA subtypes of influenza viruses but also effectively identify antigenic drift events within the same HA subtype of influenza viruses. Applications in H3N2 seasonal influenza data showed that the results from this novel method are consistent with those from the conventional serological assays. This method is not limited to the detection of antigenic variants in influenza but also other pathogens. It has the potential to be applied through a large-scale platform in disease surveillance requiring minimal biosafety and directly using clinical samples.
authors.affiliation: Department of Basic Sciences, College of Veterinary Medicine, Mississippi State University, Mississippi State, MS 39762, USA.
authors.forename: Brigitte E
authors.initials: BE
authors.lastname: Martin
authors.affiliation: Department of Basic Sciences, College of Veterinary Medicine, Mississippi State University, Mississippi State, MS 39762, USA.
authors.forename: Kun
authors.initials: K
authors.lastname: Jia
authors.affiliation: Department of Basic Sciences, College of Veterinary Medicine, Mississippi State University, Mississippi State, MS 39762, USA.
authors.forename: Hailiang
authors.initials: H
authors.lastname: Sun
authors.affiliation: Department of Basic Sciences, College of Veterinary Medicine, Mississippi State University, Mississippi State, MS 39762, USA.
authors.forename: Jianqiang
authors.initials: J
authors.lastname: Ye
authors.affiliation: Department of Basic Sciences, College of Veterinary Medicine, Mississippi State University, Mississippi State, MS 39762, USA.
authors.forename: Crystal
authors.initials: C
authors.lastname: Hall
authors.affiliation: Mississippi Public Health Laboratory, Jackson, MS 39216, USA.
authors.forename: Daphne
authors.initials: D
authors.lastname: Ware
authors.affiliation: Department of Basic Sciences, College of Veterinary Medicine, Mississippi State University, Mississippi State, MS 39762, USA. Electronic address: wan@cvm.msstate.edu.
authors.forename: Xiu-Feng
authors.initials: XF
authors.lastname: Wan
mesh_terms: D000906:Antibodies / Q000032:analysis*; D000914:Antibodies, Viral / Q000032:analysis*; D000940:Antigenic Variation; D000956:Antigens, Viral / Q000235:genetics* / Q000276:immunology; D002681:China; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000235:genetics / Q000276:immunology; D057166:High-Throughput Screening Assays / Q000295:instrumentation / Q000379:methods; D006801:Humans; D053122:Influenza A Virus, H3N2 Subtype / Q000145:classification / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D007251:Influenza, Human / Q000821:virology*; D016133:Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods*
chemical_list: D000906:Antibodies; D000914:Antibodies, Viral; D000956:Antigens, Viral; D019267:Hemagglutinin Glycoproteins, Influenza Virus
keywords: Antigenic drift; Antigenic variant; HI; Influenza virus; Neutralization