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Journal of Clinical Microbiology, February 2006, p. 291-296, Vol. 44, No. 2
0095-1137/06/$08.00+0     doi:10.1128/JCM.44.2.291-296.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Purified Excreted-Secreted Antigens from Trypanosoma cruzi Trypomastigotes as Tools for Diagnosis of Chagas' Disease

National Reference Centre for Parasitology, Montreal General Hospital, McGill University, Montreal, Québec, Canada,¹ Faculté de Médecine Vétérinaire, Université de Montréal, Montreal, Québec, Canada,² Departamento de Biología Celular, Universidad Simón Bolívar, Caracas, Venezuela,³ Ministerio de Salud y Desarrollo Social, Caracas, Venezuela⁴

Received 1 June 2005/ Returned for modification 1 August 2005/ Accepted 12 October 2005

There is currently no "gold standard" test for the diagnosis of late-stage Chagas' disease. As a result, protection of the blood supply in areas where Chagas' disease is endemic remains problematic. A panel of 709 serum samples from subjects with confirmed Chagas' disease (n = 195), healthy controls (n = 400), and patients with other parasitic diseases (n = 114) was used to assess enzyme-linked immunosorbent assays (ELISAs) based on a concentrated extract of excretory-secretory antigens from either Brazil or Tulahuen strain Trypanosoma cruzi trypomastigotes (total trypomastigote excretory-secretory antigens [TESAs]). The total TESA-based assays had excellent overall sensitivity (100%) and specificity (>94%), except for cross-reactivity with Leishmania-infected sera. In an attempt to increase the specificity of the assay, immunoaffinity chromatography was used to purify the TESA proteins (TESA_IA proteins). By Western blotting, a series of polypeptide bands with molecular masses ranging from 60 to 220 kDa were recognized by pooled sera positive for Chagas' disease. An ELISA based on TESA_IA proteins had a slightly lower sensitivity (98.6%) but an improved specificity (100%) compared to the sensitivity and specificity of the total TESA protein-based ELISAs. A 60-kDa polypeptide was identified as a major contributor to the cross-reactivity with Leishmania. These data suggest the need for field validation studies of TESA- and TESA_IA-based assays in regions where Chagas' disease is endemic.

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