Author | Andrade, Dafne Carvalho | |
Author | Borges, Igor Campos | |
Author | Laitinen, Hanna | |
Author | Ekström, Nina | |
Author | Adrian, Peter V | |
Author | Meinke, Andreas | |
Author | Barral, Aldina Maria Prado | |
Author | Carvalho, Cristiana Maria Costa Nascimento | |
Author | Käyhty, Helena | |
Access date | 2014-07-07T18:32:46Z | |
Available date | 2014-07-07T18:32:46Z | |
Document date | 2014 | |
Citation | ANDRADE, D. C. et al. A fluorescent multiplexed bead-based immunoassay (FMIA) for quantitation of IgG against Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis protein antigens. Journal of Immunological Methods, v. 405, p. 130-143, 2014. | pt_BR |
ISSN | 1872-7905 | |
URI | https://www.arca.fiocruz.br/handle/icict/7876 | |
Language | eng | pt_BR |
Publisher | Elsevier B.V | pt_BR |
Rights | open access | pt_BR |
Title | A fluorescent multiplexed bead-based immunoassay (FMIA) for quantitation of IgG against Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis protein antigens. | pt_BR |
Type | Article | pt_BR |
DOI | 10.1016/j.jim.2014.02.002 | |
Abstract | Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis are pathogens
commonly associated with infectious diseases in childhood. This study aimed to develop a
fluorescent multiplexed bead-based immunoassay (FMIA) using recombinant proteins for the
quantitation of serum IgG antibodies against these bacteria. Eight pneumococcal proteins
(Ply, CbpA, PspA1, PspA2, PcpA, PhtD, SP1732-3 and SP2216-1), 3 proteins of H. influenzae
(NTHi Protein D, NTHi0371-1, NTHi0830), and 5 proteins of M. catarrhalis (MC Omp CD,
MC_RH4_2506, MC_RH4_1701, MC_RH4_3729-1, MC_RH4_4730) were used to develop the
FMIA. Optimal coupling concentrations for each protein, comparison of singleplex and
multiplex assays, specificity, reproducibility, and correlation to ELISA for six pneumococcal
antigens were determined for validation. FMIA was then used to analyze acute and
convalescent paired serum samples of 50 children with non-severe pneumonia. The coupling
concentrations varied for different antigens, ranging from 1.6 to 32 μg of protein/million
beads. Correlation between singleplexed and multiplexed assays was excellent, with R ≥ 0.987.
The FMIA was specific, reaching N92% homologous inhibition for all specificities; heterologous
inhibition ≥20% was found only in six cases. The assay was repeatable, with averages of
intra-assay variation ≤10.5%, day-to-day variation ≤9.7% and variation between technicians
≤9.1%. Comparison with ELISA for pneumococcal antigens demonstrated good correlation with R
ranging from 0.854 (PspA2) to 0.976 (PcpA). The samples from children showed a wide range of
antibody concentrations and increases in convalescent samples. In conclusion, the FMIA was
sensitive, specific, and repeatable, using small amounts of recombinant proteins and sera to detec | pt_BR |
Affilliation | Federal University of Bahia School of Medicine. Salvador, BA, Brasil | pt_BR |
Affilliation | Federal University of Bahia School of Medicine. Salvador, BA, Brasil | pt_BR |
Affilliation | National Institute for Health and Welfare. Helsinki, Finland | pt_BR |
Affilliation | National Institute for Health and Welfare. Helsinki, Finland | pt_BR |
Affilliation | University of the Witwatersrand. DST/NRF Vaccine Preventable Diseases. Respiratory and Meningeal Pathogens Research Unit. Johannesburg, South Africa | pt_BR |
Affilliation | Valneva Austria. GmbH. Campus Vienna Biocenter. Vienna, Austria. | pt_BR |
Affilliation | Federal University of Bahia School of Medicine. Pathology Department. Salvador, BA, Brasil / Fundação Oswaldo Cruz. Centro de Pesquisa Gonçalo Moniz. Salvador, BA, Brasil. | pt_BR |
Affilliation | Federal University of Bahia School of Medicine. Department of Pediatrics. Salvador, BA, Brasil. | pt_BR |
Affilliation | National Institute for Health and Welfare. Helsinki, Finland. | pt_BR |
Subject | Antibody | pt_BR |
Subject | Multiplex | pt_BR |
Subject | Etiological diagnosis | pt_BR |
Subject | Recombinant proteins | pt_BR |
Subject | Serology | pt_BR |
Subject | Validation | pt_BR |
DeCS | Proteínas de Bactérias/imunologia | pt_BR |
DeCS | Haemophilus influenzae/imunologia | pt_BR |
DeCS | Imunoensaio/métodos | pt_BR |
DeCS | Imunoglobulina G/imunologia | pt_BR |
DeCS | Moraxella (Branhamella) catarrhalis/imunologia | pt_BR |
DeCS | Streptococcus pneumoniae/imunologia | pt_BR |
DeCS | Doença Aguda | pt_BR |
DeCS | Anticorpos Antibacterianos/sangue | pt_BR |
DeCS | Antígenos de Bactérias/imunologia | pt_BR |
DeCS | Criança | pt_BR |
DeCS | Pré-Escolar | pt_BR |
DeCS | Infecções Comunitárias Adquiridas/sangue | pt_BR |
DeCS | Fluorescência | pt_BR |
DeCS | Humanos | pt_BR |
DeCS | Imunoglobulina G/sangue | pt_BR |
DeCS | Lactente | pt_BR |
DeCS | Microesferas | pt_BR |
DeCS | Otite Média/sangue | pt_BR |
DeCS | Pneumonia/sangue | pt_BR |
DeCS | Reprodutibilidade dos Testes | pt_BR |