title: Evaluation of multiplex assay platforms for detection of influenza hemagglutinin subtype specific antibody responses.
abstract: Influenza hemagglutination inhibition (HI) and virus microneutralization assays (MN) are widely used for seroprevalence studies. However, these assays have limited field portability and are difficult to fully automate for high throughput laboratory testing. To address these issues, three multiplex influenza subtype-specific antibody detection assays were developed using recombinant hemagglutinin antigens in combination with Chembio, Luminex®, and ForteBio®platforms. Assay sensitivity, specificity, and subtype cross-reactivity were evaluated using a panel of well characterized human sera. Compared to the traditional HI, assay sensitivity ranged from 87% to 92% and assay specificity in sera collected from unexposed persons ranged from 65% to 100% across the platforms. High assay specificity (86-100%) for A(H5N1) rHA was achieved for sera from exposed or unexposed to hetorosubtype influenza HAs. In contrast, assay specificity for A(H1N1)pdm09 rHA using sera collected from A/Vietnam/1204/2004 (H5N1) vaccinees in 2008 was low (22-30%) in all platforms. Although cross-reactivity against rHA subtype proteins was observed in each assay platform, the correct subtype specific responses were identified 78%-94% of the time when paired samples were available for analysis. These results show that high throughput and portable multiplex assays that incorporate rHA can be used to identify influenza subtype specific infections.
authors.affiliation: Influenza Division, National Center for Immunization and Respiratory Diseases, Centers for Disease Control and Prevention, Atlanta, GA 30329-4027 USA. Electronic address: hix7@cdc.gov.
authors.forename: Zhu-Nan
authors.initials: ZN
authors.lastname: Li
authors.affiliation: Battelle Memorial Institute, Columbus, OH 43201, USA.
authors.forename: Kimberly M
authors.initials: KM
authors.lastname: Weber
authors.affiliation: Battelle Memorial Institute, Columbus, OH 43201, USA.
authors.forename: Rebecca A
authors.initials: RA
authors.lastname: Limmer
authors.affiliation: Battelle Memorial Institute, Columbus, OH 43201, USA.
authors.forename: Bobbi J
authors.initials: BJ
authors.lastname: Horne
authors.affiliation: Influenza Division, National Center for Immunization and Respiratory Diseases, Centers for Disease Control and Prevention, Atlanta, GA 30329-4027 USA.
authors.forename: James
authors.initials: J
authors.lastname: Stevens
authors.affiliation: Battelle Memorial Institute, Columbus, OH 43201, USA.
authors.forename: Joy
authors.initials: J
authors.lastname: Schwerzmann
authors.affiliation: Emory Vaccine Center, Emory University, Atlanta, GA 30322, USA.
authors.forename: Jens
authors.initials: J
authors.lastname: Wrammert
authors.affiliation: Emory Vaccine Center, Emory University, Atlanta, GA 30322, USA.
authors.forename: Megan
authors.initials: M
authors.lastname: McCausland
authors.affiliation: Battelle Memorial Institute, Columbus, OH 43201, USA.
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Phipps
authors.affiliation: Influenza Division, National Center for Immunization and Respiratory Diseases, Centers for Disease Control and Prevention, Atlanta, GA 30329-4027 USA.
authors.forename: Kathy
authors.initials: K
authors.lastname: Hancock
authors.affiliation: Influenza Division, National Center for Immunization and Respiratory Diseases, Centers for Disease Control and Prevention, Atlanta, GA 30329-4027 USA.
authors.forename: Daniel B
authors.initials: DB
authors.lastname: Jernigan
authors.affiliation: Influenza Division, National Center for Immunization and Respiratory Diseases, Centers for Disease Control and Prevention, Atlanta, GA 30329-4027 USA.
authors.forename: Min
authors.initials: M
authors.lastname: Levine
authors.affiliation: Influenza Division, National Center for Immunization and Respiratory Diseases, Centers for Disease Control and Prevention, Atlanta, GA 30329-4027 USA. Electronic address: jmk9@cdc.gov.
authors.forename: Jacqueline M
authors.initials: JM
authors.lastname: Katz
authors.affiliation: Influenza Division, National Center for Immunization and Respiratory Diseases, Centers for Disease Control and Prevention, Atlanta, GA 30329-4027 USA.
authors.forename: Joseph D
authors.initials: JD
authors.lastname: Miller
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000914:Antibodies, Viral / Q000097:blood*; D002648:Child; D002675:Child, Preschool; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000276:immunology*; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D007251:Influenza, Human / Q000453:epidemiology* / Q000821:virology*; D008875:Middle Aged; D012680:Sensitivity and Specificity; D012698:Serologic Tests / Q000379:methods*; D055815:Young Adult
chemical_list: D000914:Antibodies, Viral; D019267:Hemagglutinin Glycoproteins, Influenza Virus
keywords: Antibody; Antibody biosensor assay; Chembio; Hemagglutinin; Influenza; Luminex(®)