Use of Recombinant Antigens for the Accurate Immunodiagnosis of Chagas' Disease

Marco A. KriegerFundacao Oswaldo Cruz, Departamentos de Bioquimica e Biologia Molecular, Biomanguinhos, Medicina Tropical, e Genetica, Fundacao Hemocentro, Rio de Janeiro, Brazil

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Elza AlmeidaFundacao Oswaldo Cruz, Departamentos de Bioquimica e Biologia Molecular, Biomanguinhos, Medicina Tropical, e Genetica, Fundacao Hemocentro, Rio de Janeiro, Brazil

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Walter OelemannFundacao Oswaldo Cruz, Departamentos de Bioquimica e Biologia Molecular, Biomanguinhos, Medicina Tropical, e Genetica, Fundacao Hemocentro, Rio de Janeiro, Brazil

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Juan J. LafailleFundacao Oswaldo Cruz, Departamentos de Bioquimica e Biologia Molecular, Biomanguinhos, Medicina Tropical, e Genetica, Fundacao Hemocentro, Rio de Janeiro, Brazil

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Jose Borges PereiraFundacao Oswaldo Cruz, Departamentos de Bioquimica e Biologia Molecular, Biomanguinhos, Medicina Tropical, e Genetica, Fundacao Hemocentro, Rio de Janeiro, Brazil

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Henrique KriegerFundacao Oswaldo Cruz, Departamentos de Bioquimica e Biologia Molecular, Biomanguinhos, Medicina Tropical, e Genetica, Fundacao Hemocentro, Rio de Janeiro, Brazil

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Maria Ruth CarvalhoFundacao Oswaldo Cruz, Departamentos de Bioquimica e Biologia Molecular, Biomanguinhos, Medicina Tropical, e Genetica, Fundacao Hemocentro, Rio de Janeiro, Brazil

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Samuel GoldenbergFundacao Oswaldo Cruz, Departamentos de Bioquimica e Biologia Molecular, Biomanguinhos, Medicina Tropical, e Genetica, Fundacao Hemocentro, Rio de Janeiro, Brazil

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We tested two Trypanosoma cruzi recombinant antigens in a diagnostic test for Chagas' disease. These antigens were a cytoplasmic repetitive antigen (CRA) and a flagellar repetitive antigen (FRA). The results indicate that the recombinant antigens give better results when used in combination than when used separately, and that the removal of the β-galactosidase portion of the recombinant fusion proteins increases the specificity of the diagnostic test for Chagas' disease. In addition, a direct enzyme-linked immunosorbent assay (ELISA), which involves the use of peroxidase-labeled antigens to detect the immune-complexes, was developed and compared with a conventional ELISA. The results indicate that the recombinant (CRA + FRA) ELISA is better than the conventional ELISA in the diagnosis of Chagas' disease, providing 100% specificity and sensitivity in all sera tested to date. The recombinant ELISA was compared with conventional serologic tests (hemagglutination and immunofluorescence) for Chagas' disease diagnosis, and the results show that the recombinant ELISA does not give rise to false-positive results that are observed with the two other tests. The use of the recombinant ELISA should be useful in the prevention of transmission of Chagas' disease by blood transfusions.

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