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Journal of Clinical Microbiology, May 1999, p. 1554-1560, Vol. 37, No. 5
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Evaluation of Recombinant Antigens for Serodiagnosis of Chagas' Disease in South and Central America

Eufrosina S. Umezawa,1,* Sueli F. Bastos,1 Mario E. Camargo,1 Luci M. Yamauchi,2 Márcia R. Santos,2 Antonio Gonzalez,3 Bianca Zingales,4 Mariano J. Levin,5 Octavio Sousa,6 Rafael Rangel-Aldao,7 and José Franco da Silveira2

Instituto de Medicina Tropical de São Paulo, FMUSP,1 Departamento de Micro, Imuno e Parasitologia da Escola Paulista de Medicina, UNIFESP,2 and Instituto de Química, USP,4 São Paulo, Brazil; Instituto de Parasitologia y Biomedicina, CSIC, Granada, Spain3; Instituto de Investigaciones en Engenieria Genética y Biologia Molecular, Buenos Aires, Argentina5; CIDEP, Universidad de Panama, Panama6; and Universidad Simon Bolivar, Caracas, Venezuela7

Received 26 October 1998/Returned for modification 5 January 1999/Accepted 2 February 1999

The commercially available diagnostic tests for Chagas' disease employ whole extracts or semipurified fractions of Trypanosoma cruzi epimastigotes. Considerable variation in the reproducibility and reliability of these tests has been reported by different research laboratories, mainly due to cross-reactivity with other pathogens and standardization of the reagents. The use of recombinant antigens for the serodiagnosis of Chagas' disease is recommended to increase the sensitivity and specificity of serological tests. Expressed in Escherichia coli, as fusion products with glutathione S-transferase, six T. cruzi recombinant antigens (H49, JL7, A13, B13, JL8, and 1F8) were evaluated in an enzyme-linked immunosorbent assay for Chagas' disease. The study was carried out with a panel of 541 serum samples of chagasic and nonchagasic patients from nine countries of Latin America (Argentina, Bolivia, Brazil, Chile, Colombia, El Salvador, Guatemala, Honduras, and Venezuela). The optimal concentration of each recombinant antigen for coating of plates was determined with the help of 125I-labelled recombinant proteins. While the specificity of the epimastigote antigen was 84% because of false positives from leishmaniasis cases, for the recombinant antigens it varied from 96.2 to 99.6%. Recombinant antigens reacted with 79 to 100% of serum samples from chronic chagasic patients. In this way, it is proposed that a mixture of a few T. cruzi recombinant antigens should be employed in a diagnostic kit to minimize individual variation and promote high sensitivity in the diagnosis of Chagas' disease.

* Corresponding author. Mailing address: Instituto de Medicina Tropical de São Paulo, FMUSP, Av. Dr. Enéas de Carvalho Aguiar 470, CEP 05403-000, São Paulo, Brazil. Phone: 55 11 30 66 70 15. Fax: 55 11 852 36 22. E-mail: eumezawa{at}usp.br.

Journal of Clinical Microbiology, May 1999, p. 1554-1560, Vol. 37, No. 5
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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