Please use this identifier to cite or link to this item: https://www.arca.fiocruz.br/handle/icict/8224
Title: Blomia tropicalis Blo t 5 and Blo t 21 recombinant allergens might confer higher specificity to serodiagnostic assays than whole mite extract.
Authors: Carvalho, Kellyanne dos Anjos
Melo Neto, Osvaldo Pompílio de
Magalhães, Franklin Barbalho
Ponte, João Carlos Marques
Felipe, Filipe Adriano Borba
Santos, Mariese Conceição Alves dos
Lima, Givaneide dos Santos
Cruz Filho, Alvaro Augusto Souza da
Pinheiro, Carina Silva
Pontes-de-Carvalho, Lain Carlos
Neves, Neuza Maria Alcântara
Affilliation: Universidade Federal da Bahia. ProAR Núcleo de Excelência em Asma. Salvador, BA, Brasil
Fundação Oswaldo Cruz. Centro de Pesquisas Aggeu Magalhães. Recife, PE, Brasil.
Associação Caruaruense de Ensino Superior. Caruaru, PE, Brasil.
Universidade Federal da Bahia. Instituto de Ciências da Saúde. Laboratório de Alergia e Acarologia. Salvador, BA, Brasil.
Universidade Federal da Bahia. Instituto de Ciências da Saúde. Laboratório de Alergia e Acarologia. Salvador, BA, Brasil
Universidade Federal da Bahia. Instituto de Matemática. Salvador, BA, Brasil.
Centro Estudos de Alergias Respiratórias. Salvador, BA, Brasil.
Centro Estudos de Alergias Respiratórias. Salvador, BA, Brasil / Universidade Federal da Bahia. ProAR Núcleo de Excelência em Asma. Salvador, BA, Brasil.
Universidade Federal da Bahia. ProAR Núcleo de Excelência em Asma. Salvador, BA, Brasil.
Fundação Oswaldo Cruz. Centro de Pesquisa Gonçalo Moniz. Salvador, BA, Brasil.
Universidade Federal da Bahia. ProAR Núcleo de Excelência em Asma. Salvador, BA, Brasil.
Abstract: BACKGROUND: Blomia tropicalis is a dust mite and an important source of allergens in tropical regions. Up to now, the assays to diagnose atopy to this mite use whole body extract as antigens. However, anti-B. tropicalis IgE antibodies cross-react with Ascaris lumbricoides antigens, hindering the diagnosis of allergy to this mite. In this study, B. tropicalis recombinant allergens were evaluated with the purpose of developing an immunodiagnostic assay for allergy to this mite with greater specificity than those commercially available. METHODS: Two B. tropicalis allergens (Blo t 5 and Blo t 21) were cloned into a plasmidial expression vector, expressed in Escherichia coli and purified by affinity chromatography. Sixty-three sera containing anti-B. tropicalis extract (BtE) IgE antibodies were used to investigate IgE reactivity to the recombinant Blot 5 and 21 allergens. Inhibition assays with 20 sera pre-adsorbed with A. lumbricoides extract were performed using rBlo t 5, rBlo t 21, and BtE as antigens. All the assays were carried using indirect ELISA. RESULTS: Eighty-two point nine percent and 80.0% of the sera with anti-BtE antibodies from 35 children reacted with rBlo t 5 and rBlo t 21, respectively, whereas 92.8% and 89.3% of the 28 sera with anti-BtE antibodies from adult asthma patients reacted with the same allergens, and 96.4% of these sera reacted with a mixture of rBlo t 5 and rBlo t 21. In an inhibition ELISA, the absorption of sera by A. lumbricoides extract affected less the reaction with rBlo t 5 and rBlo t 21 than with BtE. CONCLUSIONS: The rBlo t 5 and rBlo t 21 allergens contain important epitopes recognized by IgE antibodies of individuals allergic to B. tropicalis antigens. Moreover, the assays using the recombinant allergens had lower IgE cross-reactivity with A. lumbricoides antigens, a fact which would confers higher specificity to serodiagnostic assays than the crude mite extract. However, additional recombinant allergens should be evaluated in order to reach the same sensitivity of the commercially available assays based on mite extract
Keywords: Blomia tropicalis
Recombinant allergens
Immunodiagnosis
Cross-reactivity
Ascaris lumbricoides
Sensitivity
Specificity
DeCS: Alérgenos/imunologia
Ascaris/imunologia
Misturas Complexas/imunologia
Ácaros/imunologia
Proteínas Recombinantes de Fusão/imunologia
Testes Sorológicos/métodos
Adulto
Animais
Antígenos de Helmintos/imunologia
Pré-Escolar
Reações Cruzadas/imunologia
Humanos
Imunoglobulina E/imunologia
Proteínas Recombinantes de Fusão/isolamento & purificação
Reprodutibilidade dos Testes
Sensibilidade e Especificidade
Issue Date: 2013
Publisher: BioMed Central
Citation: CARVALHO, K. dos A. et al. Blomia tropicalis Blo t 5 and Blo t 21 recombinant allergens might confer higher specificity to serodiagnostic assays than whole mite extract. BMC Immunology, v. 14, p.11, 2013.
DOI: 10.1186/1471-2172-14-11
ISSN: 1471-2172
Copyright: open access
Appears in Collections:BA - IGM - Artigos de Periódicos

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