title: Interlaboratory standardization of the sandwich enzyme-linked immunosorbent assay designed for MATS, a rapid, reproducible method for estimating the strain coverage of investigational vaccines.
abstract: The meningococcal antigen typing system (MATS) sandwich enzyme-linked immunosorbent assay (ELISA) was designed to measure the immunologic cross-reactivity and quantity of antigens in target strains of a pathogen. It was first used to measure the factor H-binding protein (fHbp), neisserial adhesin A (NadA), and neisserial heparin-binding antigen (NHBA) content of serogroup B meningococcal (MenB) isolates relative to a reference strain, or "relative potency" (RP). With the PorA genotype, the RPs were then used to assess strain coverage by 4CMenB, a multicomponent MenB vaccine. In preliminary studies, MATS accurately predicted killing in the serum bactericidal assay using human complement, an accepted correlate of protection for meningococcal vaccines. A study across seven laboratories assessed the reproducibility of RPs for fHbp, NadA, and NHBA and established qualification parameters for new laboratories. RPs were determined in replicate for 17 MenB reference strains at laboratories A to G. The reproducibility of RPs among laboratories and against consensus values across laboratories was evaluated using a mixed-model analysis of variance. Interlaboratory agreement was very good; the Pearson correlation coefficients, coefficients of accuracy, and concordance correlation coefficients exceeded 99%. The summary measures of reproducibility, expressed as between-laboratory coefficients of variation, were 7.85% (fHbp), 16.51% (NadA), and 12.60% (NHBA). The overall within-laboratory measures of variation adjusted for strain and laboratory were 19.8% (fHbp), 28.8% (NHBA), and 38.3% (NadA). The MATS ELISA was successfully transferred to six laboratories, and a further laboratory was successfully qualified.
authors.affiliation: Division of Bacterial Diseases, National Center for Immunization and Respiratory Diseases, Centers for Diseases Control and Prevention, Atlanta, Georgia, USA.
authors.forename: Brian D
authors.initials: BD
authors.lastname: Plikaytis
authors.forename: Maria
authors.initials: M
authors.lastname: Stella
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Boccadifuoco
authors.forename: Lisa M
authors.initials: LM
authors.lastname: DeTora
authors.forename: Mauro
authors.initials: M
authors.lastname: Agnusdei
authors.forename: Laura
authors.initials: L
authors.lastname: Santini
authors.forename: Brunella
authors.initials: B
authors.lastname: Brunelli
authors.forename: Luca
authors.initials: L
authors.lastname: Orlandi
authors.forename: Isabella
authors.initials: I
authors.lastname: Simmini
authors.forename: Marzia
authors.initials: M
authors.lastname: Giuliani
authors.forename: Morgan
authors.initials: M
authors.lastname: Ledroit
authors.forename: Eva
authors.initials: E
authors.lastname: Hong
authors.forename: Muhamed-Kheir
authors.initials: MK
authors.lastname: Taha
authors.forename: Kim
authors.initials: K
authors.lastname: Ellie
authors.forename: Gowrisankar
authors.initials: G
authors.lastname: Rajam
authors.forename: George M
authors.initials: GM
authors.lastname: Carlone
authors.forename: Heike
authors.initials: H
authors.lastname: Claus
authors.forename: Ulrich
authors.initials: U
authors.lastname: Vogel
authors.forename: Ray
authors.initials: R
authors.lastname: Borrow
authors.forename: Jamie
authors.initials: J
authors.lastname: Findlow
authors.forename: Stefanie
authors.initials: S
authors.lastname: Gilchrist
authors.forename: Paola
authors.initials: P
authors.lastname: Stefanelli
authors.forename: Cecilia
authors.initials: C
authors.lastname: Fazio
authors.forename: Anna
authors.initials: A
authors.lastname: Carannante
authors.forename: Jan
authors.initials: J
authors.lastname: Oksnes
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Fritzsønn
authors.forename: Anne-Marie
authors.initials: AM
authors.lastname: Klem
authors.forename: Dominique A
authors.initials: DA
authors.lastname: Caugant
authors.forename: Raquel
authors.initials: R
authors.lastname: Abad
authors.forename: Julio A
authors.initials: JA
authors.lastname: Vázquez
authors.forename: Rino
authors.initials: R
authors.lastname: Rappuoli
authors.forename: Mariagrazia
authors.initials: M
authors.lastname: Pizza
authors.forename: John J
authors.initials: JJ
authors.lastname: Donnelly
authors.forename: Duccio
authors.initials: D
authors.lastname: Medini
mesh_terms: D018829:Adhesins, Bacterial / Q000276:immunology; D000942:Antigens, Bacterial / Q000276:immunology; D001426:Bacterial Proteins / Q000276:immunology; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods* / Q000592:standards*; D005838:Genotype; D006801:Humans; D008585:Meningitis, Meningococcal / Q000175:diagnosis / Q000276:immunology* / Q000517:prevention & control; D022401:Meningococcal Vaccines / Q000276:immunology*; D009345:Neisseria meningitidis / Q000145:classification / Q000235:genetics / Q000276:immunology*; D018272:Porins / Q000276:immunology; D011485:Protein Binding
chemical_list: C570015:4CMenB vaccine; D018829:Adhesins, Bacterial; D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D022401:Meningococcal Vaccines; D018272:Porins; C484699:factor H-binding protein, Neisseria meningitidis; C057699:porin protein, Neisseria