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Immuno-enzymatic evaluation of the recombinant TSSA-II protein of Trypanosoma cruzi in dogs and human sera: a tool for epidemiological studies

Published online by Cambridge University Press:  26 April 2011

R. O. CIMINO ,
M. MONJE RUMI ,
P. RAGONE ,
J. LAUTHIER ,
A. ALBERTI D'AMATO ,
I. R. LÓPEZ QUIROGA ,
J. F. GIL ,
S. P. CAJAL ,
N. ACOSTA  and
M. JUÁREZ
...Show all authors
R. O. CIMINO*
Affiliation:
Cátedra de Química Biológica, Facultad de Ciencias Naturales. Universidad Nacional de Salta, Av. Bolivia 5150, Salta (4400), Argentina Instituto de Investigaciones de Enfermedades Tropicales, Sede Regional Orán, Universidad Nacional de Salta, Alvarado 751 Orán-Salta (4530), Argentina
M. MONJE RUMI
Affiliation:
Unidad de Epidemiología Molecular-Instituto de Patología Experimental, Facultad de Ciencias de la Salud. Universidad Nacional de Salta, Av. Bolivia 5150, Salta (4400), Argentina
P. RAGONE
Affiliation:
Unidad de Epidemiología Molecular-Instituto de Patología Experimental, Facultad de Ciencias de la Salud. Universidad Nacional de Salta, Av. Bolivia 5150, Salta (4400), Argentina
J. LAUTHIER
Affiliation:
Unidad de Epidemiología Molecular-Instituto de Patología Experimental, Facultad de Ciencias de la Salud. Universidad Nacional de Salta, Av. Bolivia 5150, Salta (4400), Argentina
A. ALBERTI D'AMATO
Affiliation:
Unidad de Epidemiología Molecular-Instituto de Patología Experimental, Facultad de Ciencias de la Salud. Universidad Nacional de Salta, Av. Bolivia 5150, Salta (4400), Argentina
I. R. LÓPEZ QUIROGA
Affiliation:
Cátedra de Química Biológica, Facultad de Ciencias Naturales. Universidad Nacional de Salta, Av. Bolivia 5150, Salta (4400), Argentina
J. F. GIL
Affiliation:
Cátedra de Química Biológica, Facultad de Ciencias Naturales. Universidad Nacional de Salta, Av. Bolivia 5150, Salta (4400), Argentina Instituto de Investigaciones de Enfermedades Tropicales, Sede Regional Orán, Universidad Nacional de Salta, Alvarado 751 Orán-Salta (4530), Argentina Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Argentina
S. P. CAJAL
Affiliation:
Instituto de Investigaciones de Enfermedades Tropicales, Sede Regional Orán, Universidad Nacional de Salta, Alvarado 751 Orán-Salta (4530), Argentina
N. ACOSTA
Affiliation:
Instituto de Investigaciones de Enfermedades Tropicales, Sede Regional Orán, Universidad Nacional de Salta, Alvarado 751 Orán-Salta (4530), Argentina
M. JUÁREZ
Affiliation:
Instituto de Investigaciones de Enfermedades Tropicales, Sede Regional Orán, Universidad Nacional de Salta, Alvarado 751 Orán-Salta (4530), Argentina
A. KROLEWIECKI
Affiliation:
Instituto de Investigaciones de Enfermedades Tropicales, Sede Regional Orán, Universidad Nacional de Salta, Alvarado 751 Orán-Salta (4530), Argentina
V. ORELLANA
Affiliation:
Cátedra de Química Biológica, Facultad de Ciencias Naturales. Universidad Nacional de Salta, Av. Bolivia 5150, Salta (4400), Argentina
R. ZACCA
Affiliation:
Cátedra de Microbiología, Facultad de Ciencias de la Salud de la Universidad Nacional de Salta, Av. Bolivia 5150, Salta (4400), Argentina
I. MARCIPAR
Affiliation:
Laboratorio de Tecnología Inmunológica, Universidad Nacional del Litoral, Santa Fe, Ciudad Universitaria, Paraje El pozo, CC 242 (3000) Santa Fe, Argentina
P. DIOSQUE
Affiliation:
Unidad de Epidemiología Molecular-Instituto de Patología Experimental, Facultad de Ciencias de la Salud. Universidad Nacional de Salta, Av. Bolivia 5150, Salta (4400), Argentina Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Argentina
J. R. NASSER
Affiliation:
Cátedra de Química Biológica, Facultad de Ciencias Naturales. Universidad Nacional de Salta, Av. Bolivia 5150, Salta (4400), Argentina Instituto de Investigaciones de Enfermedades Tropicales, Sede Regional Orán, Universidad Nacional de Salta, Alvarado 751 Orán-Salta (4530), Argentina
*
*Corresponding author: Cátedra de Química Biológica, Facultad de Ciencias Naturales. Universidad Nacional de Salta, Av. Bolivia 5150, Salta (4400), Argentina. E-mail: rcimino@unsa.edu.ar
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Summary

The rTSSA-II (recombinant Trypomastigote Small Surface II) antigen was evaluated by ELISA to detect anti-Trypanosoma cruzi antibodies in sera from naturally infected dogs and humans. For this evaluation ELISA-rTSSA-II was standardized and groups were classified according to the results obtained through xenodiagnosis, ELISA and PCR. Sensitivity (Se), Specificity (Sp), Kappa index (KI) and area under curve (AUC) were determined. The Se was determined by using 14 sera from dogs infected with T. cruzi VI (TcVI) whereas Sp was determined by using 95 non-chagasic sera by xenodiagnosis, ELISA-Homogenate and PCR. The performance of ELISA-rTSSA-II in dog sera was high (AUC=0·93 and KI=0·91). The Se was 92·85% (1 false negative) and Sp was 100%. Two sera from dogs infected with TcI and 1 with TcIII were negative. For patients infected with T. cruzi, reactivity was 87·8% (36/41), there was only 1 indeterminate, and Sp was 100%. Fifty-four sera from non-chagasic and 68 sera from patients with cutaneous leishmaniasis did not react with rTSS-II. ELISA-rTSSA-II showed a high performance when studying sera from naturally infected dogs and it also presented 100% Sp. This assay could be an important tool to carry out sero-epidemiological surveys on the prevalence of T. cruzi circulating lineages in the region.

Keywords

Trypanosoma cruziChagas' diseaselineageELISATSSA-IIdiagnosisArgentina
Type
Research Article
Information
Parasitology , Volume 138 , Issue 8 , July 2011 , pp. 995 - 1002
Copyright
Copyright © Cambridge University Press 2011

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References

REFERENCES

Affranchino, J. L., Ibáñez, C. F., Luquetti, R. A., Reyes, M. B., Macina, R. A., Åslund, L., Pettersson, U. and Frasch, A. (1989). Identification of a Trypanosoma cruzi antigen that is shed during the acute phase of Chagas' disease. Molecular and Biochemical Parasitology 34, 221–228.CrossRefGoogle ScholarPubMed
Aguirre, S., Silber, A. M., Brito, M. E. F., Ribone, M. E., Lagier, C. M. and Marcipar, I. S. (2006). Design, construction and evaluation of a specific chimeric antigen to diagnose Chagasic infection. Journal of Clinical Microbiology 44, 3768–3774.CrossRefGoogle ScholarPubMed
Bhattacharyya, T., Brooks, J., Yeo, M., Carrasco, H., Lewis, M., Llewellyn, M. and Miles, M. (2010). Analysis of molecular diversity of the Trypanosome cruzi trypomastigote small surface antigen reveals novel epitopes, evidence of positive selection and potential implications for lineage-specific serology. International Journal for Parasitology 40, 921–928.CrossRefGoogle ScholarPubMed
Bradford, M. M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72, 248–245.CrossRefGoogle ScholarPubMed
Britto, C., Cardoso, M., Ravel, C., Santoro, A., Pereira, J. B., Coura, J. R., Morel, C. M. and Wincker, P. (1995). Trypanosoma cruzi: parasite detection and strain discrimination in chronic chagasic patients from northeastern Brazil using PCR amplification of kinetoplast DNA and nonradioactive hybridization. Experimental Parasitology 81, 462–471.CrossRefGoogle ScholarPubMed
Burgos, J. M., Begher, S., Valadares Silva, H. M., Bisio, M., Duffy, T., Levin, M., Macedo, A. M. and Schijman, A. G. (2008). Case Report: Molecular identification of Trypanosma cruzi I tropism for central nervous system in Chagas reactivation due to AIDS. The American Journal of Tropical Medicine and Hygiene 78, 294–297.CrossRefGoogle Scholar
Buscaglia, C. and Di Noia, J. (2003). Trypanosoma cruzi clonal diversity and the epidemiology of Chagas' disease. Microbes and Infection 5, 419–427.CrossRefGoogle ScholarPubMed
Chiaramonte, M. G., Zwirner, N. W., Caropresi, S. L., Taranto, N. J. and Malchiodi, E. L. (1996). Trypanosoma cruzi and Leishmania spp. Human mixed infection. The American Journal of Tropical Medicine and Hygiene 54, 271–273.CrossRefGoogle ScholarPubMed
Caballero, Z., Sousa, O., Marques, W. P., Saez-Alquezar, A. and Umezawa, E. S. (2007). Evaluation of serological tests to identify Trypanosoma cruzi infection in humans and determine cross-reactivity with Trypanosma rangeli and Leishmania spp. Clinical and Vaccine Immunology 14, 1045–1049.CrossRefGoogle Scholar
Camussone, C., Gonzalez, V., Belluzo, M., Pujato, N., Ribone, M. E., Lagier, C. M. and Marcipar, S. (2009). Comparison of recombinant Trypanosoma cruzi peptide mixtures versus multiepitope chimeric proteins as sensitizing antigens for immunodiagnosis. Clinical and Vaccine Immunology 16, 899–905.CrossRefGoogle ScholarPubMed
Coura, J. R. and Dias, J. C. P. (2009). Epidemiology, control and surveillance of Chagas disease – 100 years after its discovery. Memórias do Instituto Oswaldo Cruz 104 (Suppl. I), 31–40.CrossRefGoogle ScholarPubMed
da Silveira, J. E., Umezawa, E. S. and Luquetti, A. O. (2001). Chagas disease: recombinant Trypanosoma cruzi antigens for serological diagnosis. Trends in Parasitology 17, 286–291.CrossRefGoogle ScholarPubMed
Di Noia, J. M., Buscaglia, C. A., De Marchi, C. R., Almeida, I. C. and Frasch, C. C. (2002). A Trypanosoma cruzi small surface molecule provides the first immunological evidence that Chagas' disease is due to a single parasite lineage. The Journal of Experimental Medicine 195, 401–413.CrossRefGoogle ScholarPubMed
Diosque, P., Barnabé, C., Padilla, A. M., Marco, D. M., Cardozo, R. M., Cimino, R. O., Nasser, J. R., Tibayrenc, M. and Basombrío, M. A. (2003). Multilocus enzyme electrophoresis analysis of Trypanosoma cruzi isolates from a geographically restricted endemic area for Chagas' disease in Argentina. International Journal for Parasitology 33, 997–1003.CrossRefGoogle ScholarPubMed
Diosque, P., Padilla, A. M., Cimino, R. O., Cardozo, R. M., Sanchez Negrette, O., Marco, J. D., Zacca, R., Meza, C., Juarez, A., Rojo, H., Rey, R., Corrales, R. M., Nasser, J. R. and Basombrío, M. A. (2004). Chagas disease in rural areas of Chaco Province, Argentina: Epidemiologic survey in humans, reservoirs, and vectors. The American Journal of Tropical Medicine and Hygiene 71, 590–593.CrossRefGoogle ScholarPubMed
Frank, F. M., Fernández, N., Taranto, N. J., Cajal, S. P., Margni, R. A., Castro, E., Thomaz-Soccol, V. and Malchiodi, E. (2003). Characterization of human infection by Leishmania spp. in the Northwest of Argentina: immune response, double infection with Trypanosoma cruzi and species of Leishmania involved. Parasitology 126, 31–39.CrossRefGoogle ScholarPubMed
Gadelha, A. A., Verçosa, A. F. A., Lorena, V. M. B., Nakasawa, M., Carvalho, A. B., Souza, W. V., Ferreira, A. G. P., Silva, E. D., Krieger, M. A., Goldenberg, S. and Gomes, Y. M. (2003). Chagas' disease diagnosis: comparative analysis of recombinant ELISA with conventional ELISA and the haemagglutination test. Vox Sanguinis 85, 165–170.CrossRefGoogle ScholarPubMed
Gardner, I. and Greiner, M. (2006). Receiver operating characteristic curves and likelihood ratios: improvements over traditional methods for the evaluation and application of veterinary clinical pathology tests. Veterinary Clinical Pathology 35, 8–17.CrossRefGoogle ScholarPubMed
Laemmli, U. K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London 227, 680–685.CrossRefGoogle ScholarPubMed
Lauricella, M. A., Castañera, M. B., Gürtler, R. E. and Segura, E. L. (1998). Immunodiagnosis of Trypanosoma cruzi (Chagas' Disease) Infection in Naturally Infected Dogs. Memórias do Instituto Oswaldo Cruz 93, 501–507.CrossRefGoogle ScholarPubMed
Lauricella, M. A., Wisnivesky-Colli, C., Gürtler, R., Petersen, R., Bujas, M. and Segura, E. L., (1993). Standardization of serological test for detecting anti-Trypanosoma cruzi antibodies in dogs. Memórias do Instituto Oswaldo Cruz 88, 413–417.CrossRefGoogle ScholarPubMed
Minter, D. (1978). Efectos de la presencia de animales domésticos en viviendas infestadas sobre la transmisión de la enfermedad de Chagas en el Hombre. Boletín de la Oficina Sanitaria Panamerican 84, 332–343.Google Scholar
Nasser, J. R. (1997). Proteínas recombinantes del Trypanosoma cruzi con actividad inmunogénica: respuesta inmune celular y humoral. Tesis doctoral. Universidad Nacional de Tucumán. Facultad de Ciencias Naturales e Instituto Miguel Lillo San Miguel de Tucamán, Argentina.Google Scholar
Nasser, J. R., Gómez, L. E., Sánchez, D., Guerin, M. and Basombrío, M. (1997). Immunogenicity of the recombinant SAPA Protein of Trypanosoma cruzi for mice. The Journal of Parasitology 83, 76–81.CrossRefGoogle ScholarPubMed
Pereira-Chioccola, V. L., Fragata-Filho, A. A., de Apparecida Levy, A. M., Rodrigues, M. M. and Schekman, S. (2003). Enzyme-linked immunoassay using recombinant trans-sialidase of Trypanosoma cruzi can be employed for monitoring of patients with Chagas' disease drug treatment. Clinical and Diagnostic Laboratory Immunology 10, 826–830.Google Scholar
Reyes, M. B., Lorca, M., Muñoz, P. and Frasch, A. C. (1990) Fetal IgG specificities against Trypanosoma cruzi antigens in infected newborns. Proceedings of the National Academy of Science, USA 87, 2846–2850.CrossRefGoogle ScholarPubMed
Robertson, E. A. and Zweig, M. H. (1981). Use of Receiver Operating Characteristic curves to evaluate the clinical performance of analytical systems. Clinical Chemistry 27, 1569–1574.CrossRefGoogle ScholarPubMed
Sambrook, J., Fritsch, E. F. and Maniatis, T. (1989). Molecular Cloning. A Laboratory Manual. 2nd Edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA.Google Scholar
Sim, J. and Wright, C. (2005). Kappa statistic in reliability studies: use, interpretations and sample size requirements. Physical Therapy 85, 257–268.CrossRefGoogle ScholarPubMed
Umezawa, E. S., Bastos, S. F., Camargo, M. E., Yamauchi, L. M., Santos, M. R., Gonzalez, A., Zingales, B., Levin, M., Sousa, O., Rangel-Aldao, R. and da Silveira, F. (1999). Evaluation of recombinant antigens for serodiagnosis of Chagas’ disease in South and Central America. Journal of Clinical Microbiology 37, 1554–1560.CrossRefGoogle ScholarPubMed
Umezawa, E. S., Bastos, S. F., Coura, J., Levin, M., Gonzalez, A., Rangel-Aldao, R., Zingales, B., Luquetti, A. O. and da Silveira, J. F. (2003). An improved serodiagnostic test for Chagas' disease employing a mixture of Trypanosoma cruzi recombinant antigens. Transfusion 43, 91–97.CrossRefGoogle ScholarPubMed
Umezawa, E. S., Luquetti, A. O., Levitus, G., Ponce, C., Ponce, E., Henriquez, D., Revollo, S., Espinoza, B., Sousa, O., Kahn, B. and da Silveira, J. F. (2004). Serodiagnosis of chronic and acute Chagas' disease with Trypanosoma cruzi recombiant proteins: results of a collaborative study in six Latin American countries. Journal of Clinical Microbiology 42, 449–452.CrossRefGoogle Scholar
Wincker, P., Bosseno, M. F., Britto, C., Yaksic, N., Cardoso, M. A., Morel, C. M., and Brenière, S. F. (1994). High correlation between Chagas' disease serology and PCR-based detection of Trypanosoma cruzi kinetoplast DNA in Bolivian children living in an endemic area. FEMS Microbiology Letters 124, 419–423.CrossRefGoogle Scholar
Zingales, B., Andrade, S. G., Briones, M. R. S., Campbell, D. A., Chiari, E., Fernandes, O., Guhl, F., Lages-Silva, E., Macedo, A. M., Machado, C. R., Miles, M. A., Romanha, A. J., Sturm, N. R., Tibayrenc, M. and Schijman, A. G. (2009). A new consensus for Trypanosoma cruzi intraspecific nomenclature: second revision meeting recommends TcI to TcVI. Memórias do Instituto Oswaldo Cruz 104, 1051–1054.CrossRefGoogle ScholarPubMed