Trypanosoma cruzi III causing the indeterminate form of Chagas disease in a semi-arid region of Brazil Kiev Martins, Cléber de Mesquita Andrade, Andressa.

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Trypanosoma cruzi III causing the indeterminate form of Chagas disease in a semi-arid region of Brazil Kiev Martins, Cléber de Mesquita Andrade, Andressa Noronha Barbosa-Silva, Gerson Barbosa do Nascimento, Egler Chiari, Lúcia Maria da Cunha Galvão, Antonia Cláudia Jácome da Câmara International Journal of Infectious Diseases Volume 39, Pages 68-75 (October 2015) DOI: 10.1016/j.ijid.2015.08.012 Copyright © 2015 The Authors Terms and Conditions

Figure 1 Flowchart for patient classification, as recommended by the Brazilian Consensus on Chagas Disease.38 International Journal of Infectious Diseases 2015 39, 68-75DOI: (10.1016/j.ijid.2015.08.012) Copyright © 2015 The Authors Terms and Conditions

Figure 2 Protocol for genotyping proposed by D’Ávila et al.42 using a three-step assay: polymorphism analysis of the mitochondrial cytochrome oxidase subunit 2 (COII) gene, amplification of the D7 divergent domain of the 24Sα rRNA gene, and amplification of the spliced leader intergenic region (SL-IRac). International Journal of Infectious Diseases 2015 39, 68-75DOI: (10.1016/j.ijid.2015.08.012) Copyright © 2015 The Authors Terms and Conditions

Figure 3 A RFLP analysis of the mitochondrial COII gene in the Trypanosoma cruzi isolates belonging to different haplotypes, obtained by polyacrylamide gel electrophoresis. Digestion of the DNA with AluI generates three RFLP patterns for the T. cruzi strains: restriction fragments of 264, 81, and 30bp are classified as TcI (mitochondrial haplotype A; lane 1, Col1.7G2 clone), restriction fragments of 212 and 81bp are classified as TcII (mitochondrial haplotype C; lane 2, strain JG), and restriction fragments of 294 and 81bp are classified as TcIII–VI (mitochondrial haplotype B; lane 3, strain RN19; lane 4, strain AM64; lane 5, strain 3253; lane 6, CL Brener clone). Lanes 7–13 show T. cruzi samples of patients 2137, 3188, RN26, RN79, 105, CBS195, and SM76. (B) Analysis of 24Sα rRNA of T. cruzi isolates from chronic chagasic patients and controls. Lane 1, Col1.7G2 clone (∼110bp); lane 2, JG (∼125bp); lane 3, RN19 (∼110bp); lane 4, AM64 (∼117/119bp); lane 5, 3253 (∼110 and 125bp); lane 6, CL Brener clone (∼125bp); lanes 7–13, T. cruzi sample of patients 2137, 3188, RN26, RN79, 105, CBS195, and SM76. (C) The SL-IRac gene was used to separate isolates of TcIII and TcIV from other DTUs. Lanes 1, 2, 5 and 6: controls Col1.7G2 clone, JG, 3253, and CL Brener clone (fragments of 150–157bp); lanes 4 and 5: controls RN19 and AM64 (fragments of 200bp); lanes 7–13: T. cruzi sample of patients 2137, 3188, RN26, RN79, 105, CBS195, and SM76; lane M: molecular size marker; lane NC: negative PCR control. International Journal of Infectious Diseases 2015 39, 68-75DOI: (10.1016/j.ijid.2015.08.012) Copyright © 2015 The Authors Terms and Conditions

Figure 4 Distribution of discrete typing units (DTUs) of Trypanosoma cruzi in different municipalities of the State of Rio Grande do Norte, Brazil. International Journal of Infectious Diseases 2015 39, 68-75DOI: (10.1016/j.ijid.2015.08.012) Copyright © 2015 The Authors Terms and Conditions