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. 1997 Jul;35(7):1829–1834. doi: 10.1128/jcm.35.7.1829-1834.1997

Serological diagnosis of Chagas' disease: a potential confirmatory assay using preserved protein antigens of Trypanosoma cruzi.

R P Mendes 1, S Hoshino-Shimizu 1, A M Moura da Silva 1, I Mota 1, R A Heredia 1, A O Luquetti 1, P G Leser 1
PMCID: PMC229851  PMID: 9196203

Abstract

The diagnosis of Chagas' disease relies mostly on data provided by immunologic tests, but inconclusive results often require elucidation, especially in blood banks. When six different types of Trypanosoma cruzi epimastigote antigens were studied by an immunoblotting assay (IBA), a preserved protein antigen (Ag PP) was found to present the most interesting immunochemical features because of its high reactivity with anti-T. cruzi antibodies. Thus, the IBA with Ag PP (PP IBA) was assessed with panels of coded and noncoded serum samples prepared in different laboratories, including the Brazilian Reference Laboratory for Chagas' Disease. It was found that serum samples from patients proved (clinically, eletrocardiographically, serologically, and epidemiologically) to have Chagas' disease consistently recognized 12 bands (140, 100, 85, 78, 59, 57, 46, 35, 27, 23, 20, and 18 kDa) of Ag PP. In contrast, sera from nonchagasic patients, including patients with mucocutaneous leishmaniasis, were negative or reacted weakly, and one serum sample did not have more than five different bands. These bands were 78, 57, 46, 35, 27, 23, 20, or 18 kDa. A criterion was adopted to interpret the results obtained in the PP IBA. The criterion considered positive a serum sample recognizing all 12 bands and considered negative a serum sample that did not recognize any of the bands except the eight nonspecific bands mentioned above. The PP IBA indicated maximum sensitivity and specificity as well as high positive and negative predictive values. The data demonstrate that the PP IBA discriminates chagasic from nonchagasic infections and seems to be applicable as a confirmatory assay for elucidating inconclusive results obtained by standard serology.

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Selected References

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  1. Araujo F. G., Guptill D. Use of antigen preparations of the amastigote stage of Trypanosoma cruzi in the serology of Chagas' disease. Am J Trop Med Hyg. 1984 May;33(3):362–371. doi: 10.4269/ajtmh.1984.33.362. [DOI] [PubMed] [Google Scholar]
  2. Camargo M. E. Fluorescent antibody test for the serodiagnosis of American trypanosomiasis. Technical modification employing preserved culture forms of Trypanosoma cruzi in a slide test. Rev Inst Med Trop Sao Paulo. 1966 Sep-Oct;8(5):227–235. [PubMed] [Google Scholar]
  3. Camargo M. E., Hoshino S., Siqueira G. R. Hemagglutination with preserved, sensitized cells, a practical test for routine serologic diagnosis of American trypanosomiasis. Rev Inst Med Trop Sao Paulo. 1973 Mar-Apr;15(2):81–85. [PubMed] [Google Scholar]
  4. Castellani O., Ribeiro L. V., Fernandes J. F. Differentiation of Trypanosoma cruzi in culture. J Protozool. 1967 Aug;14(3):447–451. doi: 10.1111/j.1550-7408.1967.tb02024.x. [DOI] [PubMed] [Google Scholar]
  5. Chiller T. M., Samudio M. A., Zoulek G. IgG antibody reactivity with Trypanosoma cruzi and Leishmania antigens in sera of patients with Chagas' disease and leishmaniasis. Am J Trop Med Hyg. 1990 Dec;43(6):650–656. doi: 10.4269/ajtmh.1990.43.650. [DOI] [PubMed] [Google Scholar]
  6. Cotrim P. C., Paranhos G. S., Mortara R. A., Wanderley J., Rassi A., Camargo M. E., da Silveira J. F. Expression in Escherichia coli of a dominant immunogen of Trypanosoma cruzi recognized by human chagasic sera. J Clin Microbiol. 1990 Mar;28(3):519–524. doi: 10.1128/jcm.28.3.519-524.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Ferreira A. W., Belem Z. R., Moura M. E., Camargo M. E. Aspectos da padronizaço de testes sorológicos para a doença de Chagas: um teste imunoenzimático para a triagem de doadores de sangue. Rev Inst Med Trop Sao Paulo. 1991 Mar-Apr;33(2):123–128. [PubMed] [Google Scholar]
  8. Galel S. A., Kirchhoff L. V. Risk factors for Trypanosoma cruzi infection in California blood donors. Transfusion. 1996 Mar;36(3):227–231. doi: 10.1046/j.1537-2995.1996.36396182140.x. [DOI] [PubMed] [Google Scholar]
  9. Grant I. H., Gold J. W., Wittner M., Tanowitz H. B., Nathan C., Mayer K., Reich L., Wollner N., Steinherz L., Ghavimi F. Transfusion-associated acute Chagas disease acquired in the United States. Ann Intern Med. 1989 Nov 15;111(10):849–851. doi: 10.7326/0003-4819-111-10-849. [DOI] [PubMed] [Google Scholar]
  10. Hoshino-Shimizu S., Camargo M. E., Nagasse T. K. A stable polysaccharide-hemagglutination reagent for the diagnosis of acute or recent Trypanosoma cruzi infections. Rev Inst Med Trop Sao Paulo. 1978 Jul-Aug;20(4):208–212. [PubMed] [Google Scholar]
  11. Kerndt P. R., Waskin H. A., Kirchhoff L. V., Steurer F., Waterman S. H., Nelson J. M., Gellert G. A., Shulman I. A. Prevalence of antibody to Trypanosoma cruzi among blood donors in Los Angeles, California. Transfusion. 1991 Nov-Dec;31(9):814–818. doi: 10.1046/j.1537-2995.1991.31992094668.x. [DOI] [PubMed] [Google Scholar]
  12. Kirchhoff L. V., Gam A. A., Gusmao R. A., Goldsmith R. S., Rezende J. M., Rassi A. Increased specificity of serodiagnosis of Chagas' disease by detection of antibody to the 72- and 90-kilodalton glycoproteins of Trypanosoma cruzi. J Infect Dis. 1987 Mar;155(3):561–564. doi: 10.1093/infdis/155.3.561. [DOI] [PubMed] [Google Scholar]
  13. Krieger M. A., Almeida E., Oelemann W., Lafaille J. J., Pereira J. B., Krieger H., Carvalho M. R., Goldenberg S. Use of recombinant antigens for the accurate immunodiagnosis of Chagas' disease. Am J Trop Med Hyg. 1992 Apr;46(4):427–434. doi: 10.4269/ajtmh.1992.46.427. [DOI] [PubMed] [Google Scholar]
  14. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  15. Lorca M., Gonzalez A., Veloso C., Reyes V., Vergara U. Immunodetection of antibodies in sera from symptomatic and asymptomatic Chilean Chagas' disease patients with Trypanosoma cruzi recombinant antigens. Am J Trop Med Hyg. 1992 Jan;46(1):44–49. doi: 10.4269/ajtmh.1992.46.44. [DOI] [PubMed] [Google Scholar]
  16. Luquetti A. O. Use of Trypanosoma cruzi defined proteins for diagnosis--Multicentre trial. Serological and technical aspects. Mem Inst Oswaldo Cruz. 1990 Oct-Dec;85(4):497–505. doi: 10.1590/s0074-02761990000400021. [DOI] [PubMed] [Google Scholar]
  17. MAEKELT G. A. DIAGN'OSTICO DE LABORATORIO DE LAS TRIPANOSOMIASIS AMERICANAS. Rev Venez Sanid Asist Soc. 1964 Mar;29:1–18. [PubMed] [Google Scholar]
  18. Malchiodi E. L., Chiaramonte M. G., Taranto N. J., Zwirner N. W., Margni R. A. Cross-reactivity studies and differential serodiagnosis of human infections caused by Trypanosoma cruzi and Leishmania spp; use of immunoblotting and ELISA with a purified antigen (Ag163B6). Clin Exp Immunol. 1994 Sep;97(3):417–423. doi: 10.1111/j.1365-2249.1994.tb06104.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Minoprio P., Itohara S., Heusser C., Tonegawa S., Coutinho A. Immunobiology of murine T. cruzi infection: the predominance of parasite-nonspecific responses and the activation of TCRI T cells. Immunol Rev. 1989 Dec;112:183–207. doi: 10.1111/j.1600-065x.1989.tb00558.x. [DOI] [PubMed] [Google Scholar]
  20. Moncayo A., Luquetti A. O. Multicentre double blind study for evaluation of Trypanosoma cruzi defined antigens as diagnostic reagents. Mem Inst Oswaldo Cruz. 1990 Oct-Dec;85(4):489–495. doi: 10.1590/s0074-02761990000400020. [DOI] [PubMed] [Google Scholar]
  21. Morgado M. G., Ivo-dos-Santos J., Pinho R. T., Argüelles E., Rezende J. M., Galvão-Castro B. Trypanosoma cruzi: identification of specific epimastigote antigens by human immune sera. Mem Inst Oswaldo Cruz. 1989 Jul-Sep;84(3):309–314. doi: 10.1590/s0074-02761989000300004. [DOI] [PubMed] [Google Scholar]
  22. Nickerson P., Orr P., Schroeder M. L., Sekla L., Johnston J. B. Transfusion-associated Trypanosoma cruzi infection in a non-endemic area. Ann Intern Med. 1989 Nov 15;111(10):851–853. doi: 10.7326/0003-4819-111-10-851. [DOI] [PubMed] [Google Scholar]
  23. Rangel-Aldao R., Comach G., Allende O., Cayama E., Delgado V., Piras R., Piras M., Henriquez D., Negri S. Trypanosoma cruzi: polypeptide markers of epimastigotes and trypomastigotes. Mol Biochem Parasitol. 1986 Jul;20(1):25–32. doi: 10.1016/0166-6851(86)90139-8. [DOI] [PubMed] [Google Scholar]
  24. Reed S. G., Badaró R., Masur H., Carvalho E. M., Lorenco R., Lisboa A., Teixeira R., Johnson W. D., Jr, Jones T. C. Selection of a skin test antigen for American visceral leishmaniasis. Am J Trop Med Hyg. 1986 Jan;35(1):79–85. doi: 10.4269/ajtmh.1986.35.79. [DOI] [PubMed] [Google Scholar]
  25. Schechter M., Nogueira N. Variations induced by different methodologies in Trypanosoma cruzi surface antigen profiles. Mol Biochem Parasitol. 1988 May;29(1):37–45. doi: 10.1016/0166-6851(88)90117-x. [DOI] [PubMed] [Google Scholar]
  26. Schmuñis G. A. Trypanosoma cruzi, the etiologic agent of Chagas' disease: status in the blood supply in endemic and nonendemic countries. Transfusion. 1991 Jul-Aug;31(6):547–557. doi: 10.1046/j.1537-2995.1991.31691306255.x. [DOI] [PubMed] [Google Scholar]
  27. Teixeira M. G., Borges-Pereira J., Netizert E., Souza M. L., Peralta J. M. Development and evaluation of an enzyme linked immunotransfer blot technique for serodiagnosis of Chagas' disease. Trop Med Parasitol. 1994 Dec;45(4):308–312. [PubMed] [Google Scholar]
  28. Vergara U., Veloso C., Gonzalez A., Lorca M. Evaluation of an enzyme-linked immunosorbent assay for the diagnosis of Chagas' disease using synthetic peptides. Am J Trop Med Hyg. 1992 Jan;46(1):39–43. doi: 10.4269/ajtmh.1992.46.39. [DOI] [PubMed] [Google Scholar]
  29. Wong R. C., Hudson L., Hindmarsh P. J. Immune precipitation and immunoblotting for the detection of Trypanosoma cruzi antigens. Trans R Soc Trop Med Hyg. 1986;80(2):275–281. doi: 10.1016/0035-9203(86)90034-9. [DOI] [PubMed] [Google Scholar]

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