TY - JOUR
T1 - Interlaboratory standardization of the sandwich enzyme-linked immunosorbent assay designed for MATS, a rapid, reproducible method for estimating the strain coverage of investigational vaccines
AU - Plikaytis, Brian D.
AU - Stella, Maria
AU - Boccadifuoco, Giuseppe
AU - DeTora, Lisa M.
AU - Agnusdei, Mauro
AU - Santini, Laura
AU - Brunelli, Brunella
AU - Orlandi, Luca
AU - Simmini, Isabella
AU - Giuliani, Marzia
AU - Ledroit, Morgan
AU - Hong, Eva
AU - Taha, Muhamed Kheir
AU - Ellie, Kim
AU - Rajam, Gowrisankar
AU - Carlone, George M.
AU - Claus, Heike
AU - Vogel, Ulrich
AU - Borrow, Ray
AU - Findlow, Jamie
AU - Gilchrist, Stefanie
AU - Stefanelli, Paola
AU - Fazio, Cecilia
AU - Carannante, Anna
AU - Oksnes, Jan
AU - Fritzsønn, Elisabeth
AU - Klem, Anne Marie
AU - Caugan, Dominique A.
AU - Abad, Raquel
AU - Vázquez, Julio A.
AU - Rappuoli, Rino
AU - Pizza, Mariagrazia
AU - Donnelly, John J.
AU - Medini, Duccio
PY - 2012/10
Y1 - 2012/10
N2 - 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 withinlaboratory 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.
AB - 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 withinlaboratory 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.
UR - http://www.scopus.com/inward/record.url?scp=84867281306&partnerID=8YFLogxK
U2 - 10.1128/CVI.00202-12
DO - 10.1128/CVI.00202-12
M3 - Article
C2 - 22875603
AN - SCOPUS:84867281306
SN - 1556-6811
VL - 19
SP - 1609
EP - 1617
JO - Clinical and Vaccine Immunology
JF - Clinical and Vaccine Immunology
IS - 10
ER -