Double-antigen sandwich ELISA based on chimeric antigens for detection of antibodies to Trypanosoma cruzi in human sera

dc.creatorFreitas, Natália Erdens Maron
dc.creatorSantos, Emily Ferreira
dc.creatorLeony, Leonardo Maia
dc.creatorSilva, Ângelo Antônio Oliveira
dc.creatorDaltro, Ramona Tavares
dc.creatorVasconcelos, Larissa de Carvalho Medrado
dc.creatorDuarte, Gabriela Agra
dc.creatorMota, Cristiane Oliveira da
dc.creatorSilva, Edimilson Domingos
dc.creatorCeledon, Paola Alejandra Fiorani
dc.creatorZanchin, Nilson Ivo Tonin
dc.creatorSantos, Fred Luciano Neves
dc.date2022-04-06T18:46:23Z
dc.date2022-04-06T18:46:23Z
dc.date2022
dc.date.accessioned2023-09-26T22:43:51Z
dc.date.available2023-09-26T22:43:51Z
dc.identifierFREITAS, Natália Erdens Maron et al. Double-antigen sandwich ELISA based on chimeric antigens for detection of antibodies to Trypanosoma cruzi in human sera. PLoS Neglected Tropical Diseases, V. 16, n. 3, p. 115, 2022.
dc.identifier1935-2735
dc.identifierhttps://www.arca.fiocruz.br/handle/icict/52068
dc.identifier10.1371/journal.pntd.0010290
dc.identifier.urihttps://repositorioslatinoamericanos.uchile.cl/handle/2250/8882485
dc.descriptionInstituto Gonçalo Moniz (IGM). Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES). Fundação de Amparo à Pesquisa do Estado da Bahia (FAPESB). Prêmio Inova Fiocruz. Prêmio Estudos e Projetos (FINEP). Banco Nacional de Desenvolvimento Prêmio Econômico e Social (BNDES).
dc.descriptionBackground: Enzyme-linked immunosorbent assays (ELISA) are generally the chosen test for Chagas disease (CD) diagnosis; however, its performance depends on the antigen preparation adsorbed to the solid phase, which may lead to false-positive results and cross-reactions. The use of chimeric recombinant antigens can overcome this limitation. Four chimeric antigens from Trypanosoma cruzi (IBMP-8.1, IBMP-8.2, IBMP-8.3 and IBMP-8.4) were developed and evaluated in phase I, II and III studies using indirect ELISA as diagnostic platform. However, peroxidase-labeled secondary anti-human IgG antibody, which is employed in indirect ELISAs, limits its use for the detection of species-specific and class-specific antibodies. To overcome this limitation, peroxidase-labeled antigens can be utilized, diagnosing both acute or chronic infection, in a species and immunoglobulin class-independent manner, through the use of a double-antigen sandwich ELISA (DAgS-ELISA). We aimed to evaluate and validate the diagnostic performance of the chimeric antigens IBMP-8.1, IBMP-8.2, IBMP-8.3 and IBMP-8.4 in the DAgS-ELISA platform. Methodology/Principal findings: DAgS-ELISA was optimized by checkerboard titration. In phase I study, 207 positive and 205 negative samples were evaluated. Cross-reactivity to other infections was also assessed using 68 samples. The selected conditions for the tests utilized 25 ng of antigen per well and the conjugate diluted at 1:2,000 for all molecules. In the phase I study, the areas under the curve of IBMP-8.1, IBMP-8.2, IBMP-8.3 and IBMP-8.4 were 98.7%, 99.5%, 98.6% and 98.8%, respectively. Among the positive samples, IBMP-8.1 antigen classified 53 (25.6%) as false negative, IBMP-8.2, 27 (13%), IBMP-8.3, 24 (11.6%) and IBMP-8.4, 43 (20.8%), giving sensitivities of 74.4%, 87%, 88.4% and 79.2%, respectively. The only antigen that did not reach 100% specificity was IBMP-8.3, with 96.6%. IBMP-8.3 was also the only molecule to show cross-reactivity with HTLV. Conclusions/Significance: DAgS-ELISA is a promising tool for immunodiagnosis, and despite the high AUC values, the performance of this assay was different from the values obtained by our group when using these antigens in the indirect ELISA, for this reason, improvements are being considered to increase the sensitivity of the DAgS-ELISA.
dc.formatapplication/pdf
dc.languageeng
dc.publisherPublic Library of Science
dc.rightsopen access
dc.subjectTrypanosoma cruzi
dc.subjectEnsaio de Imunoadsorção Enzimática
dc.subjectAnticorpos
dc.subjectTrypanosoma cruzi
dc.subjectEnzyme-Linked Immunosorbent Assay
dc.subjectAntibodies
dc.subjectTrypanosoma cruzi
dc.subjectEnsayo de Inmunoadsorción Enzimática
dc.subjectAnticuerpos
dc.subjectTrypanosoma cruzi
dc.subjectTest ELISA
dc.subjectAnticorps
dc.titleDouble-antigen sandwich ELISA based on chimeric antigens for detection of antibodies to Trypanosoma cruzi in human sera
dc.typeArticle


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