Identification of Trypanosoma cruzi Discrete Typing Units (DTUs) in Latin-American migrants in Barcelona (Spain). Issue 2 (April 2017)

• Record Type: Journal Article
• Authors: Abras, Alba; Gállego, Montserrat; Muñoz, Carmen; Juiz, Natalia A; Ramírez, Juan Carlos; Cura, Carolina I; Tebar, Silvia; Fernández-Arévalo, Anna; Pinazo, María-Jesús; de la Torre, Leonardo; Posada, Elizabeth; Navarro, Ferran; Espinal, Paula; Ballart, Cristina; Portús, Montserrat; Gascón, Joaquim; Schijman, Alejandro G
• Is Part Of: Parasitology international. Volume 66:Issue 2(2017:Apr.)
• Page Start: 83
• Page End: 88
• Publication Date: 2017-04
• Subjects: Chagas disease -- Trypanosoma cruzi -- Discrete Typing Unit -- Migration -- Real-time PCR -- Sequencing; Parasitology -- Periodicals; Parasites -- Periodicals; Parasitic Diseases -- Periodicals; Parasitology -- Periodicals; Parasitologie -- Périodiques; 571.99905
• DOI: 10.1016/j.parint.2016.12.003
• Languages: English
• ISSNs: 1383-5769

Abstract

Abstract: Trypanosoma cruzi, the causative agent of Chagas disease, is divided into six Discrete Typing Units (DTUs): TcI–TcVI. We aimed to identify T. cruzi DTUs in Latin-American migrants in the Barcelona area (Spain) and to assess different molecular typing approaches for the characterization of T. cruzi genotypes. Seventy-five peripheral blood samples were analyzed by two real-time PCR methods (qPCR) based on satellite DNA (SatDNA) and kinetoplastid DNA (kDNA). The 20 samples testing positive in both methods, all belonging to Bolivian individuals, were submitted to DTU characterization using two PCR-based flowcharts: multiplex qPCR using TaqMan probes (MTq-PCR), and conventional PCR. These samples were also studied by sequencing the SatDNA and classified as type I (TcI/III), type II (TcII/IV) and type I/II hybrid (TcV/VI). Ten out of the 20 samples gave positive results in the flowcharts: TcV (5 samples), TcII/V/VI (3) and mixed infections by TcV plus TcII (1) and TcV plus TcII/VI (1). By SatDNA sequencing, we classified the 20 samples, 19 as type I/II and one as type I. The most frequent DTU identified by both flowcharts, and suggested by SatDNA sequencing in the remaining samples with low parasitic loads, TcV, is common in Bolivia and predominant in peripheral blood. The mixed infection by TcV–TcII was detected for the first time simultaneously in Bolivian migrants. PCR-based flowcharts are very useful to characterize DTUs during acute infection. SatDNA sequence analysis cannot discriminate T. cruzi populations at the level of a single DTU but it enabled us to increase the number of characterized cases in chronically infected patients. Graphical abstract: Highlights: PCR flowcharts and satellite-DNA sequencing were used to genotype Trypanosoma cruzi . Parasitic load of chronic patients was limiting for flowcharts but not for satellite-DNA sequencing. Most DTUs identified belonged to samples from neonates and pediatric cases. The most identified DTU was TcV, common in Bolivian domestic cycles. A mixed infection TcV–TcII was detected for the first time simultaneously in Bolivian migrants.