Use of two recombinant proteins of Trypanosoma cruzi in the serological diagnosis of Chagas disease

Zingales, Bianca; Gruber, Arthur; Ramalho, C. B.; Umezawa, Eufrosina Setsu; Colli, Walter

doi:10.1590/s0074-02761990000400024

Cited by 13 publications

(12 citation statements)

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“…Moreover, our specificity estimate of ϳ90% for samples from leishmaniasis patients is likely to be an overestimate since no samples from patients with visceral leishmaniasis were included in our panel. Our data suggest that the concentrated total TESAs from both the Brazil and the Tulahuen strains include conserved epitopes that are highly immunogenic, which meets one of the principal criteria for Chagas' disease tests suggested by the World Health Organization (32).…”

Section: Discussionmentioning

confidence: 56%

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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Section: Discussionmentioning

confidence: 56%

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

et al. 2006

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“…Several research groups have therefore used recombinant/synthetic and biochemically purified antigens (Affranchino et al 1989, Goldenberg et al 1991, Krieger et al 1992, Umezawa et al 1999, Franco da Silveira et al 2001. In order to be useful, these antigens must meet several criteria: (i) they should be present in T. cruzi isolates from different endemic areas, and absent from other infectious disease agents; (ii) they should be highly immunogenic in populations with different genetic backgrounds, irrespective of the clinical phase of Chagas disease and (iii) they should be stable and easily amenable to qualitycontrol tests, to guarantee reproducibility (Zingales et al 1990, Stolf 1992). …”

Section: What Still Needs To Be Done Regarding Diagnosis?mentioning

confidence: 99%

Diagnosis of Chagas disease: what has been achieved? What remains to be done with regard to diagnosis and follow up studies?

Gomes

Lorena

Luquetti

2009

Mem. Inst. Oswaldo Cruz

122

109

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“…Several T. cruzi genes were isolated and cloned, and some recombinant antigens have been assayed and tested for diagnosis. The results, however, showed variable efficacy for the different tested antigens, and it was observed that some of them displayed better performance when used in combination than separately, producing results with sensitivities and specificities that reached up to 100% (1,2,17,23,24,31,32,40,49,54).…”

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confidence: 99%

Trypanosoma cruziRecombinant Complement Regulatory Protein: a Novel Antigen for Use in an Enzyme-Linked Immunosorbent Assay for Diagnosis of Chagas' Disease

Meira¹,

Galvão²,

Gontijo

et al. 2002

J Clin Microbiol

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Currently, diagnosis of Chagas' disease is based on serological methods, but due to the high occurrence of inconclusive results, more reliable methods are needed. The use of recombinant antigens for serodiagnosis of Chagas' disease is recommended in order to increase the sensitivity and specificity of the serological tests. The Trypanosoma cruzi complement regulatory protein (CRP) is a surface glycoprotein present on the trypomastigote forms of the parasite, and the recombinant CRP (rCRP) was cloned in a mammalian expression system and purified by affinity chromatography. The purified recombinant protein was used as an antigen in an enzyme-linked immunosorbent assay (rCRP ELISA) in order to verify its sensitivity and specificity compared with other established methods. In this evaluation, a panel of 184 serum samples distributed among chronic chagasic patients (n ‫؍‬ 65), blood bank donors (n ‫؍‬ 100), and patients infected with Leishmania spp. (n ‫؍‬ 19) was used. The sensitivity and specificity of the rCRP ELISA were 100% when compared to conventional serology and complement-mediated lysis tests from these groups. When hemoculture and PCR tests were evaluated for diagnosis of chronic chagasic patients, using the rCRP ELISA as a reference test, the positivities were found to be 64.62 and 81.54%, respectively, showing a higher degree of sensitivity of the test. The data demonstrate that rCRP ELISA was able to discriminate between chronic chagasic patients and nonchagasic individuals, such as blood donors and patients with leishmaniasis. Thus, the rCRP is an excellent antigen for use in Chagas' disease diagnosis, due to the absence of false-negative or false-positive results.

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Use of two recombinant proteins of Trypanosoma cruzi in the serological diagnosis of Chagas disease

Cited by 13 publications

References 0 publications

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

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

Diagnosis of Chagas disease: what has been achieved? What remains to be done with regard to diagnosis and follow up studies?

Trypanosoma cruziRecombinant Complement Regulatory Protein: a Novel Antigen for Use in an Enzyme-Linked Immunosorbent Assay for Diagnosis of Chagas' Disease

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