Identification of a Western Blot Pattern for the Specific Diagnosis of Trypanosoma cruzi Infection in Human Sera

Cristina Riera Laboratori de Parasitologia, Facultat de Farmàcia, Universitat de Barcelona, Barcelona, Spain

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Mireia Verges Laboratori de Parasitologia, Facultat de Farmàcia, Universitat de Barcelona, Barcelona, Spain

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Laura Iniesta Laboratori de Parasitologia, Facultat de Farmàcia, Universitat de Barcelona, Barcelona, Spain

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Roser Fisa Laboratori de Parasitologia, Facultat de Farmàcia, Universitat de Barcelona, Barcelona, Spain

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Montserrat Gállego Laboratori de Parasitologia, Facultat de Farmàcia, Universitat de Barcelona, Barcelona, Spain

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Silvia Tebar Laboratori de Parasitologia, Facultat de Farmàcia, Universitat de Barcelona, Barcelona, Spain

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Montserrat Portús Laboratori de Parasitologia, Facultat de Farmàcia, Universitat de Barcelona, Barcelona, Spain

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A Western blot (WB) method using a lysate from Trypanosoma cruzi (Maracay strain) epimastigotes was evaluated. Serum samples from 37 patients with confirmed Chagas disease (cohort I), 27 Spanish patients with visceral leishmaniasis caused by Leishmania infantum (cohort II), and 28 Colombian patients with cutaneous leishmaniasis caused by L. panamensis and negative serology for Chagas disease (cohort III) were tested. The negative controls were 55 healthy seronegative subjects for T. cruzi and Leishmania; 28 of the negative controls were from a region endemic for Chagas disease and Leishmania (cohort IV), and 27 of the negative controls were from a non-endemic area for Leishmania and T. cruzi (cohort V). A homogeneous standard band pattern consisting of six antigenic bands corresponding to 28, 32, 38, 39, 40, and 48 kDa was recognized simultaneously for all Chagasic patients' sera. Sera from Leishmania-infected patients showed a heterogeneous band pattern that was easily differentiated from the pattern of patients with Chagas disease. WB with T. cruzi epimastigote antigen is an efficient method for diagnosis and may be used as an alternative to confirm T. cruzi and detect cross-reactivity with Leishmania.

Author Notes

*Address correspondence to Cristina Riera, Laboratori de Parasitologia, Facultat de Farmàcia, Universitat de Barcelona, Avda. Joan XXIII s.n., E-08028 Barcelona, Spain. E-mail: mcriera@ub.edu

Authors' addresses: Cristina Riera, Mireia Verges, Laura Iniesta, Roser Fisa, Montserrat Gállego, Silvia Tebar, and Montserrat Portús, Laboratori de Parasitologia, Facultat de Farmàcia, Universitat de Barcelona, Barcelona, Spain, E-mails: mcriera@ub.edu, mireiaverges@hotmail.com, laura.iniesta@gmail.com, silviatebar@ub.edu, rfisa@ub.edu, mgallego@ub.edu, and mportus@ub.edu.

Reprint requests: Cristina Riera, Laboratori de Parasitologia, Facultat de Farmàcia, Universitat de Barcelona, Avda. Joan XXIII s.n., E-08028 Barcelona, Spain, E-mail: mcriera@ub.edu.

  • 1.

    World Health Organization, 2007. Guhl F, Lazdins-Helds JK, eds. Reporte del grupo de trabajo cientiìfico sobre la enfermedad de Chagas. Geneva, Switzerland: World Health Organization.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 2.

    Schmunis GA, 2007. Epidemiology of Chagas disease in non-endemic countries: the role of international migration. Mem Inst Oswaldo Cruz 102: 7585.

  • 3.

    Schmunis GA, Yadon ZE, 2010. Chagas disease: a Latin American health problem becoming a world health problem. Acta Trop 115: 1421.

  • 4.

    Riera C, Guarro A, Kassab HE, Jorba JM, Castro M, Angrill R, Gállego M, Fisa R, Martin C, Lobato A, Portús M, 2006. Congenital transmission of Trypanosoma cruzi in Europe (Spain): a case report. Am J Trop Med Hyg 75: 10781081.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 5.

    Centers for Disease Control and Prevention (CDC), 2006. Chagas disease after organ transplantation—Los Angeles, California, 2006. MMWR Morb Mortal Wkly Rep 55: 798800.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 6.

    Muñoz J, Portús M, Corachán M, Fumadó V, Gascón J, 2007. Congenital Trypanosoma cruzi infection in a non-endemic area. Trans R Soc Trop Med Hyg 101: 11611162.

  • 7.

    Young C, Losikoff P, Chawla A, Glasser L, Forman E, 2007. Transfusion-acquired Trypanosoma cruzi infection. Transfusion 47: 540544.

  • 8.

    Flores-Chávez M, Faez Y, Olalla JM, Cruz I, Gárate T, Rodríguez M, Blanc P, Cañavate C, 2008. Fatal congenital Chagas' disease in a non-endemic area: a case report. Cases J 1: 302.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 9.

    Chang CD, Cheng KY, Jiang LX, Salbilla VA, Haller AS, Yem AW, Bryant JD, Kirchhoff LV, Leiby DA, Schochetman G, Shah DO, 2006. Evaluation of a prototype Trypanosoma cruzi antibody assay with recombinant antigens on a fully automated chemiluminescence analyzer for blood donor screening. Transfusion 46: 17371744.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 10.

    World Health Organization Expert Committee, 2002. Control of Chagas Disease. World Health Organization Technical Report Series 905. Geneva, Switzerland: World Health Organization, 1109.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 11.

    Frade AF, Luquetti AO, Prata A, Ferreira AW, 2011. Western blotting method (TESAcruzi) as a supplemental test for confirming the presence of anti-Trypanosoma cruzi antibodies in finger prick blood samples from children aged 0–5 years in Brazil. Acta Trop 117: 1013.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 12.

    Reiche EM, Cavazzana M Jr, Okamura H, Tagata EC, Jankevicius SI, Jankevicius JV, 1998. Evaluation of the western blot in the confirmatory serologic diagnosis of Chagas' disease. Am J Trop Med Hyg 59: 750756.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 13.

    Flores-Chávez M, Fernández B, Puente S, Torres P, Rodríguez M, Monedero C, Cruz I, Gárate T, Cañavate C, 2008. Transfusional Chagas disease: parasitological and serological monitoring of an infected recipient and blood donor. Clin Infect Dis 46: 4447.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 14.

    Riera C, Vergés M, López-Chejade P, Piron M, Gascón J, Fisa R, Gállego M, Portús M, 2009. Desarrollo y evaluación de una técnica ELISA con antígeno crudo de Trypanosoma cruzi para el diagnóstico de la Enfermedad de Chagas. Enf Emerg 11: 2229.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 15.

    Riera C, Valladares JE, Gállego M, Aisa MJ, Castillejo S, Fisa R, Ribas N, Carrió J, Alberola J, Arboix M, 1999. Serological and parasitological follow-up in dogs experimentally infected with Leishmania infantum and treated with meglumine antimoniate. Vet Parasitol 84: 3347.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 16.

    Aisa MJ, Castillejo S, Gallego M, Fisa R, Riera MC, de Colmenares M, Torras S, Roura X, Sentis J, Portús M, 1998. Diagnostic potential of Western blot analysis of sera from dogs with leishmaniasis in endemic areas and significance of the pattern. Am J Trop Med Hyg 58: 154159.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 17.

    Malchiodi EL, Chiaramonte MG, Taranto NJ, Zwirner NW, 1994. 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 97: 417423.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 18.

    Umezawa EF, Nascimento NS, Kesper N Jr, Coura JR, Borges-Pereira J, Junqueira AC, Camargo ME, 1996. Immunoblot assay using excreted-secreted antigen of Trypanosoma cruzi in serodiagnosis of congenital, acute, and chronic Chagas disease. J Clin Microbiol 34: 21432147.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 19.

    Oelemann WM, Vanderborght BO, Verissimo Da Costa GC, Teixeira MG, Borges-Pereira J, De Castro JA, Coura JR, Stoops E, Hulstaert F, Zrein M, Peralta JM, 1999. A recombinant peptide antigen line immunoassay optimized for the confirmation of Chagas'  disease. Transfusion 39: 711717.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 20.

    Bucio MI, Cabrera M, Segura EL, Zenteno E, Salazar-Schettino M, 1999. Identification of immunodominant antigens in Mexican strains of Trypanosoma cruzi. Immunol Invest 28: 257268.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 21.

    Sánchez B, Monteon V, Reyes PA, Espinoza B, 2001. Standardization of micro-enzyme-linked immunosorbent assay (ELISA) and Western blot for detection of Trypanosoma cruzi antibodies using extracts from Mexican strains as antigens. Arch Med Res 32: 382388.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 22.

    Berrizbeitia M, Ndao M, Bubis J, Gottschalk M, Aché A, Lacouture S, Medina M, Ward BJ, 2006. Purified excreted-secreted antigens from Trypanosona cruzi trypomastigotes as tools for diagnosis of Chagas' disease. J Clin Microbiol 44: 291296.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 23.

    Campos Y, Briceño L, Reina K, Figarella K, Pérez JL, Mosca W, 2009. Serological diagnosis of Chagas disease: evaluation and characterisation of a low cost antigen with high sensitivity and specificity. Mem Inst Oswaldo Cruz 104: 914917.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 24.

    Teixeira MG, Borges-Pereira J, Netizert E, Souza ML, Peralta JM, 1994. Development and evaluation of an enzyme linked immunotransfer blot technique for serodiagnosis of Chagas' disease. Trop Med Parasitol 45: 308312.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 25.

    Marcipar IS, Risso MG, Silber AM, Revelli S, Marcipar AJ, 2001. Antibody maturation in Trypanosoma cruzi-infected rats. Clin Diagn Lab Immunol 8: 802805.

  • 26.

    Camargo ME, Rebonato C, 1969. Cross-reactivity in fluorescence tests for Trypanosoma and Leishmania antibodies. A simple inhibition procedure to ensure specific results. Am J Trop Med Hyg 18: 500505.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 27.

    Guimarães MC, Celeste BJ, de Castilho EA, Mineo JR, Diniz JM, 1981. Immunoenzymatic assay (ELISA) in mucocutaneous leishmaniasis, kala-azar, and Chagas'  disease: an epimastigote Trypanosoma cruzi antigen able to distinguish between anti-Trypanosoma and anti-Leishmania antibodies. Am J Trop Med Hyg 30: 942947.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 28.

    Araujo FG, 1986. Analysis of Trypanosoma cruzi antigens bound by specific antibodies and by antibodies to related trypanosomatids. Infect Immun 53: 179185.

  • 29.

    Dos Santos JI, Morgado MG, Galvão-Castro B, 1987. Human visceral leishmaniasis: analysis of the specificity of humoral immune response to polypeptides of Leishmania donovani chagasi. Am J Trop Med Hyg 37: 263270.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 30.

    Evans TG, Krug EC, Wilson ME, Vasconcelos AW, de Alencar JE, Pearson RD, 1989. Evaluation of antibody responses in American visceral leishmaniasis by ELISA and immunoblot. Mem Inst Oswaldo Cruz 84: 157166.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 31.

    Vexenat Ade C, Santana JM, Teixeira AR, 1996. Cross-reactivity of antibodies in human infections by the kinetoplastid protozoa Trypanosoma cruzi, Leishmania chagasi and Leishmania (Viannia) braziliensis. Rev Inst Med Trop Sao Paulo 38: 177185.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 32.

    Gonçalves CC, Reiche EM, De Abreu Filho BA, Silveira TG, Felizardo TC, Maia KR, Costacurta R, Padovesi EJ, Dias Filho BP, Jankevicius SI, Jankevicius JV, 2002. Evaluation of antigens from various Leishmania species in a Western blot for diagnosis of American tegumentary leishmaniasis. Am J Trop Med Hyg 66: 91102.

    • PubMed
    • Search Google Scholar
    • Export Citation
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