El ensayo inmunoenzimatico en microgotas sobre nitrocelulosa (Dot-ELISA) en el diagnostico de la enfermedad de Chagas: II. Estudio seroepidemiologico en cuatro comunidades rurales de Venezuela

SUMMARY. Many serological tests have been developed for the diagnosis of ChagasÕ disease, but few have been subjected to a rigorous field evaluation. We have recently described several novel enzyme immunoassays (EIAs) based on fixed-whole organisms or trypomastigote excretory-secretory antigens (TESA) from different Trypanosoma cruzi strains (Tulahuen or Brazil). This study evaluated the most promising of these novel assays (e.g. fixed-epimastigotes, fixedtrypomastigotes, TESA Brazil and TESA Tulahuen antigens) in a field study of Venezuelan blood bank specimens. The assays were tested in an operatorblinded fashion using 2038 blood bank samples obtained from low and high T. cruzi prevalence regions of Venezuela (n ¼ 1050 and n ¼ 988 from Bolivar and Portuguesa states, respectively). Based on National Laboratory for Chagas Immunodiagnosis (NLCI)Ôgold standardÕ results, all novel EIAs were superior to the commercial kit currently used in Venezuela, achieving 100% sensitivity and >99% specificity at optimal cut-off values. The novel assays identified seven false-negative samples compared with the routine screening performed by the Venezuelan blood bank although two samples were also misclassified as positive. Minor differences in the performance of the four novel assays were observed at lower arbitrary cutoff values. This study confirms the potential utility of both the fixed-organism and the TESA-based assays in the diagnosis of T. cruzi infection.

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Field evaluation of four novel enzyme immunoassays for Chagas’ disease in Venezuela blood banks: comparison of assays using fixed‐epimastigotes, fixed‐trypomastigotes or trypomastigote excreted–secreted antigens from two Trypanosoma cruzi strains

Berrizbeitia

Ndao

Bubis

et al. 2006

Transfusion Medicine

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Development and Comparison of Enzyme Immunoassays for Diagnosis of Chagas' Disease Using Fixed Forms of Trypanosoma cruzi (Epimastigotes, Amastigotes, and Trypomastigotes) and Assessment of Antigen Stability for the Three Assays

et al. 2004

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Three enzyme immunoassays (EIAs) for diagnosis of Chagas' disease were developed with fixed forms of Trypanosoma cruzi using a panel of 435 sera from the following groups: Venezuelan subjects positive by immunofluorescence (n ‫؍‬ 70), Venezuelan healthy controls (n ‫؍‬ 85), healthy Canadians (n ‫؍‬ 166), and subjects with other parasitic diseases (n ‫؍‬ 114). All assays achieved 100% sensitivity and reasonable specificity for amastigotes (97.6%), epimastigotes (98.3%), and trypomastigotes (99.3%). The fixed-trypomastigote assay was stable over 4 months at 4°C and room temperature. These data suggest that a fixed-trypomastigote EIA may be a suitable candidate for blood bank screening.

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

et al. 2006

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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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El ensayo inmunoenzimatico en microgotas sobre nitrocelulosa (Dot-ELISA) en el diagnostico de la enfermedad de Chagas: II. Estudio seroepidemiologico en cuatro comunidades rurales de Venezuela

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Field evaluation of four novel enzyme immunoassays for Chagas’ disease in Venezuela blood banks: comparison of assays using fixed‐epimastigotes, fixed‐trypomastigotes or trypomastigote excreted–secreted antigens from two Trypanosoma cruzi strains

Field evaluation of four novel enzyme immunoassays for Chagas’ disease in Venezuela blood banks: comparison of assays using fixed‐epimastigotes, fixed‐trypomastigotes or trypomastigote excreted–secreted antigens from two Trypanosoma cruzi strains

Development and Comparison of Enzyme Immunoassays for Diagnosis of Chagas' Disease Using Fixed Forms of Trypanosoma cruzi (Epimastigotes, Amastigotes, and Trypomastigotes) and Assessment of Antigen Stability for the Three Assays

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

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