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      Detection of anti- Trypanosoma cruzi antibodies by chimeric antigens in chronic Chagas disease-individuals from endemic South American countries

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          Abstract

          Background

          Laboratory diagnosis of chronic Chagas disease is a troubling factor due to lack of reference tests. The WHO suggests the use of two distinct commercial serological tests in parallel. The performance of commercial immunoassays might fluctuate depending on the antigenic matrices and the local strains of T. cruzi in different geographical settings. The use of antigenic matrices based on chimeric proteins can solve these limitations. Here, we evaluated the diagnostic performance of two chimeric T. cruzi antigens (IBMP-8.1 and -8.4) to diagnose chronic Chagas disease in individuals from endemic South American countries.

          Methodology/Principal findings

          IBMP-8.1 and IBMP-8.4 chimeric antigens were expressed as soluble proteins in E. coli and purified using chromatography methods. Reactivity of IBMP-8.1 and IBMP-8.4 was assessed using an in-house ELISA with sera from 122 non-infected and 215 T. cruzi-infected individuals from Argentina, Bolivia, and Paraguay. Cut-off values were based on ROC curves and performance parameters were determined using a dichotomous approach. Area under the curve values were > 99.7% for both IBMP-8.1 and IBMP-8.4 antigens. IgG levels in T. cruzi-positive and negative samples were higher for IBMP-8.4 than IBMP-8.1. Both IBMP-8.1 and -8.4 were 100% specific, while IBMP-8.4 were 100% sensitive compared to IBMP-8.1 (95.3%). Admitting RI values of 1.0 ± 0.10 as the inconclusive interval, 6.2% of the samples tested using IBMP-8.1 and 2.1% using IBMP-8.4 fell inside the grey zone. Based on accuracy and diagnostic odds ratio values, IBMP-8.4 presented the best performance. Differences in sensitivity and IgG levels among the samples from Argentina, Bolivia, and Paraguay were not significant.

          Conclusions/Significance

          Our findings showed a notable performance of IBMP-8.1 and -8.4 chimeric antigens in diagnosing chronic Chagas disease in individuals from endemic South American countries, confirming our hypothesis that these antigens could be used in geographical areas where distinct T. cruzi DTUs occur.

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          Most cited references22

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          The revised Trypanosoma cruzi subspecific nomenclature: rationale, epidemiological relevance and research applications.

          The protozoan Trypanosoma cruzi, its mammalian reservoirs, and vectors have existed in nature for millions of years. The human infection, named Chagas disease, is a major public health problem for Latin America. T. cruzi is genetically highly diverse and the understanding of the population structure of this parasite is critical because of the links to transmission cycles and disease. At present, T. cruzi is partitioned into six discrete typing units (DTUs), TcI-TcVI. Here we focus on the current status of taxonomy-related areas such as population structure, phylogeographical and eco-epidemiological features, and the correlation of DTU with natural and experimental infection. We also summarize methods for DTU genotyping, available for widespread use in endemic areas. For the immediate future multilocus sequence typing is likely to be the gold standard for population studies. We conclude that greater advances in our knowledge on pathogenic and epidemiological features of these parasites are expected in the coming decade through the comparative analysis of the genomes from isolates of various DTUs. Copyright © 2012 Elsevier B.V. All rights reserved.
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            Clinical and epidemiological aspects of Chagas disease.

            A. Prata (2001)
            Chagas disease is caused by the protozoan parasite Trypanosoma cruzi. During the past decades, after urban migrations, Chagas disease became frequent in cities and a health problem in non-endemic countries, where it can be transmitted vertically and by blood transfusion or organ transplantation. Microepidemics of acute Chagas disease have been reported, probably due to oral transmission. Heart involvement is the major feature of the disease because of its characteristics, frequency, and consequences, and is also the source of most controversies. The indeterminate clinical form, despite its good prognosis on at least a medium-term basis (5-10 years), has acquired increasing importance due to the controversial meaning of the abnormality of some tests and the myocardial focal lesions found in many patients. Simultaneous evaluation of the parasympathetic and of the sympathetic system in the heart has been done by spectral analysis of heart rate. The physiopathological and clinical significance of denervation in Chagas disease is still incompletely understood. There are major divergences of opinion on specific treatment during the chronic phase because of the doubts about cure rates. Changes of Chagas disease prevalence in many countries have been certified by the Pan American Health Organization, and are ascribed to large-scale vector-control programmes with modern pyrethroid insecticides and to improvement in lifestyle.
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              Trypanosoma cruzi genetic diversity: Something new for something known about Chagas disease manifestations, serodiagnosis and drug sensitivity.

              The genetic diversity of Trypanosoma cruzi, the protozoan agent of Chagas disease, is widely recognized. At present, T. cruzi is partitioned into seven discrete typing units (DTUs), TcI-TcVI and Tcbat. This article reviews the present knowledge on the parasite population structure, the evolutionary relationships among DTUs and their distinct, but not exclusive ecological and epidemiological associations. Different models for the origin of hybrid DTUs are examined, which agree that genetic exchange among T. cruzi populations is frequent and has contributed to the present parasite population structure. The geographic distribution of the prevalent DTUs in humans from the southern United States to Argentina is here presented and the circumstantial evidence of a possible association between T. cruzi genotype and Chagas disease manifestations is discussed. The available information suggests that parasite strains detected in patients, regardless of the clinical presentation, reflect the principal DTU circulating in the domestic transmission cycles of a particular region. In contrast, in several orally transmitted outbreaks, sylvatic strains are implicated. As a consequence of the genotypic and phenotypic differences of T. cruzi strains and the differential geographic distribution of DTUs in humans, regional variations in the sensitivity of the serological tests are verified. The natural resistance to benznidazole and nifurtimox, verified in vivo and in vitro for some parasite stocks, is not associated with any particular DTU, and does not explain the marked difference in the anti-parasitic efficacy of both drugs in the acute and chronic phases of Chagas disease. Throughout this review, it is emphasized that the interplay between parasite and host genetics should have an important role in the definition of Chagas disease pathogenesis, anti-T. cruzi immune response and chemotherapy outcome and should be considered in future investigations.
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                Author and article information

                Contributors
                Role: Formal analysisRole: MethodologyRole: ValidationRole: Writing – original draftRole: Writing – review & editing
                Role: ValidationRole: Writing – original draftRole: Writing – review & editing
                Role: InvestigationRole: MethodologyRole: Writing – review & editing
                Role: Funding acquisitionRole: InvestigationRole: MethodologyRole: Writing – review & editing
                Role: ResourcesRole: Writing – review & editing
                Role: Project administrationRole: ValidationRole: Writing – review & editing
                Role: InvestigationRole: ResourcesRole: Writing – review & editing
                Role: ConceptualizationRole: Data curationRole: Funding acquisitionRole: InvestigationRole: Writing – original draftRole: Writing – review & editing
                Role: ConceptualizationRole: Data curationRole: Formal analysisRole: InvestigationRole: MethodologyRole: Project administrationRole: SupervisionRole: ValidationRole: VisualizationRole: Writing – original draftRole: Writing – review & editing
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                18 April 2019
                2019
                : 14
                : 4
                : e0215623
                Affiliations
                [1 ] Faculty of Technology and Sciences of Bahia, Salvador, Bahia, Brazil
                [2 ] Gonçalo Moniz Institute, Oswaldo Cruz Foundation, Salvador, Bahia, Brazil
                [3 ] Molecular Biology Institute of Paraná, Curitiba, Paraná, Brazil
                [4 ] Carlos Chagas Institute, Oswaldo Cruz Foundation, Curitiba, Paraná, Brazil
                [5 ] Department of Pathology and Legal Medicine, Federal University of Bahia, Salvador, Bahia, Brazil
                [6 ] Department of Epidemiology of Microbial Diseases, School of Public Health, Yale University, New Haven, Connecticut, United States of America
                [7 ] Aggeu Magalhães Institute, Oswaldo Cruz Foundation, Recife, Pernambuco, Brazil
                [8 ] Laboratory of Molecular Biology of Chagas Disease, Institute for Research on Genetic Engineering and Molecular Biology “Dr Héctor Torres”, Buenos Aires, Argentina
                Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, BRAZIL
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                Author information
                http://orcid.org/0000-0002-3944-0818
                Article
                PONE-D-19-09603
                10.1371/journal.pone.0215623
                6472793
                30998741
                d4975e8e-2fde-4c4b-927f-c7b675bff17f
                © 2019 Del-Rei et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 8 January 2019
                : 4 April 2019
                Page count
                Figures: 4, Tables: 0, Pages: 12
                Funding
                Funded by: funder-id http://dx.doi.org/10.13039/501100008777, Fundación Bunge y Born;
                Award ID: 2012
                Award Recipient :
                Funded by: Consejo Nacional de Investigaciones Científicas y Tecnológicas
                Award ID: PIP/0974-2011
                Award Recipient :
                Funded by: Gonçalo Moniz Institute
                Award ID: PROEP/IGM 400904/2013-6
                Award Recipient :
                Funded by: Coordination of Superior Level Staff Improvement
                Award ID: CAPES - PROEX 0720/2018
                Award Recipient :
                Funded by: National Council for Scientific and Technological Development
                Award ID: CNPq Proc. No. 312195/2015-0
                Award Recipient :
                Funded by: National Council for Scientific and Technological Development
                Award ID: CNPq Proc. No. 307319/2016-4
                Award Recipient :
                This work was supported by: Fundación Bunge y Born 2012 Dr. Silvia Andrea Longhi; Consejo Nacional de Investigaciones Científicas y Tecnológicas PIP/0974-2011 Dr. Alejandro Gabriel Schijman; Gonçalo Moniz Institute PROEP/IGM 400904/2013-6 Dr. Mitermayer Galvão dos Reis; Coordination of Superior Level Staff Improvement CAPES - PROEX 0720/2018 Dr. Fred Luciano Neves Santos; National Council for Scientific and Technological Development CNPq Proc. No. 312195/2015-0 Dr. Nilson Ivo Tonin Zanchin; and National Council for Scientific and Technological Development CNPq Proc. No. 307319/2016-4 Dr. Mitermayer Galvão dos Reis. The funders were not involved in the design of the study, in the collection, analysis and interpretation of the data, or in writing the manuscript.
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