title: Identification of Trypanosoma cruzi Discrete Typing Units (DTUs) in Latin-American migrants in Barcelona (Spain).
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.
authors.affiliation: Secció de Parasitologia, Departament de Biologia, Sanitat i Medi Ambient, Facultat de Farmàcia, Universitat de Barcelona, Av. Joan XXIII 27-31, 08028 Barcelona, Spain; ISGlobal, Barcelona Centre for International Health Research (CRESIB), Roselló 134-4°, 08036 Barcelona, Spain; Laboratori d'Ictiologia Genètica, Departament de Biologia, Universitat de Girona, Campus de Montilivi, 17071 Girona, Spain.
authors.forename: Alba
authors.initials: A
authors.lastname: Abras
authors.affiliation: Secció de Parasitologia, Departament de Biologia, Sanitat i Medi Ambient, Facultat de Farmàcia, Universitat de Barcelona, Av. Joan XXIII 27-31, 08028 Barcelona, Spain; ISGlobal, Barcelona Centre for International Health Research (CRESIB), Roselló 134-4°, 08036 Barcelona, Spain. Electronic address: mgallego@ub.edu.
authors.forename: Montserrat
authors.initials: M
authors.lastname: Gállego
authors.affiliation: Servei de Microbiologia, Hospital de la Santa Creu i Sant Pau, Sant Quintí 89, 08041 Barcelona, Spain; Institut d'Investigació Biomèdica Sant Pau (IIB Sant Pau), Hospital de la Santa Creu i Sant Pau, Sant Antoni Maria Claret 167, 08025 Barcelona, Spain; Departament de Genètica i Microbiologia, Universitat Autònoma de Barcelona, Campus de Bellaterra, 08193 Cerdanyola del Vallès, Spain.
authors.forename: Carmen
authors.initials: C
authors.lastname: Muñoz
authors.affiliation: Laboratorio de Biología Molecular de la Enfermedad de Chagas (LaBMECh), Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres" (INGEBI-CONICET), Vuelta de Obligado 2490-2°, C1428ADN Ciudad Autónoma de Buenos Aires, Argentina.
authors.forename: Natalia A
authors.initials: NA
authors.lastname: Juiz
authors.affiliation: Laboratorio de Biología Molecular de la Enfermedad de Chagas (LaBMECh), Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres" (INGEBI-CONICET), Vuelta de Obligado 2490-2°, C1428ADN Ciudad Autónoma de Buenos Aires, Argentina.
authors.forename: Juan Carlos
authors.initials: JC
authors.lastname: Ramírez
authors.affiliation: Laboratorio de Biología Molecular de la Enfermedad de Chagas (LaBMECh), Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres" (INGEBI-CONICET), Vuelta de Obligado 2490-2°, C1428ADN Ciudad Autónoma de Buenos Aires, Argentina.
authors.forename: Carolina I
authors.initials: CI
authors.lastname: Cura
authors.affiliation: Secció de Parasitologia, Departament de Biologia, Sanitat i Medi Ambient, Facultat de Farmàcia, Universitat de Barcelona, Av. Joan XXIII 27-31, 08028 Barcelona, Spain; ISGlobal, Barcelona Centre for International Health Research (CRESIB), Roselló 134-4°, 08036 Barcelona, Spain.
authors.forename: Silvia
authors.initials: S
authors.lastname: Tebar
authors.affiliation: Secció de Parasitologia, Departament de Biologia, Sanitat i Medi Ambient, Facultat de Farmàcia, Universitat de Barcelona, Av. Joan XXIII 27-31, 08028 Barcelona, Spain; ISGlobal, Barcelona Centre for International Health Research (CRESIB), Roselló 134-4°, 08036 Barcelona, Spain; Institut d'Investigació Biomèdica Sant Pau (IIB Sant Pau), Hospital de la Santa Creu i Sant Pau, Sant Antoni Maria Claret 167, 08025 Barcelona, Spain.
authors.forename: Anna
authors.initials: A
authors.lastname: Fernández-Arévalo
authors.affiliation: ISGlobal, Barcelona Centre for International Health Research (CRESIB), Roselló 134-4°, 08036 Barcelona, Spain.
authors.forename: María-Jesús
authors.initials: MJ
authors.lastname: Pinazo
authors.affiliation: ISGlobal, Barcelona Centre for International Health Research (CRESIB), Roselló 134-4°, 08036 Barcelona, Spain.
authors.forename: Leonardo
authors.initials: L
authors.lastname: de la Torre
authors.affiliation: ISGlobal, Barcelona Centre for International Health Research (CRESIB), Roselló 134-4°, 08036 Barcelona, Spain.
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Posada
authors.affiliation: Servei de Microbiologia, Hospital de la Santa Creu i Sant Pau, Sant Quintí 89, 08041 Barcelona, Spain; Institut d'Investigació Biomèdica Sant Pau (IIB Sant Pau), Hospital de la Santa Creu i Sant Pau, Sant Antoni Maria Claret 167, 08025 Barcelona, Spain; Departament de Genètica i Microbiologia, Universitat Autònoma de Barcelona, Campus de Bellaterra, 08193 Cerdanyola del Vallès, Spain.
authors.forename: Ferran
authors.initials: F
authors.lastname: Navarro
authors.affiliation: Institut d'Investigació Biomèdica Sant Pau (IIB Sant Pau), Hospital de la Santa Creu i Sant Pau, Sant Antoni Maria Claret 167, 08025 Barcelona, Spain.
authors.forename: Paula
authors.initials: P
authors.lastname: Espinal
authors.affiliation: Secció de Parasitologia, Departament de Biologia, Sanitat i Medi Ambient, Facultat de Farmàcia, Universitat de Barcelona, Av. Joan XXIII 27-31, 08028 Barcelona, Spain; ISGlobal, Barcelona Centre for International Health Research (CRESIB), Roselló 134-4°, 08036 Barcelona, Spain.
authors.forename: Cristina
authors.initials: C
authors.lastname: Ballart
authors.affiliation: Secció de Parasitologia, Departament de Biologia, Sanitat i Medi Ambient, Facultat de Farmàcia, Universitat de Barcelona, Av. Joan XXIII 27-31, 08028 Barcelona, Spain.
authors.forename: Montserrat
authors.initials: M
authors.lastname: Portús
authors.affiliation: ISGlobal, Barcelona Centre for International Health Research (CRESIB), Roselló 134-4°, 08036 Barcelona, Spain.
authors.forename: Joaquim
authors.initials: J
authors.lastname: Gascón
authors.affiliation: Laboratorio de Biología Molecular de la Enfermedad de Chagas (LaBMECh), Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres" (INGEBI-CONICET), Vuelta de Obligado 2490-2°, C1428ADN Ciudad Autónoma de Buenos Aires, Argentina.
authors.forename: Alejandro G
authors.initials: AG
authors.lastname: Schijman
mesh_terms: D000293:Adolescent; D000328:Adult; D001838:Bolivia / Q000453:epidemiology; D014355:Chagas Disease / Q000097:blood / Q000453:epidemiology / Q000208:ethnology* / Q000469:parasitology*; D002648:Child; D060085:Coinfection / Q000453:epidemiology / Q000469:parasitology; D016054:DNA, Protozoan / Q000235:genetics*; D005260:Female; D014644:Genetic Variation; D005838:Genotype; D006801:Humans; D007231:Infant, Newborn; D008297:Male; D008875:Middle Aged; D058889:Molecular Typing; D059208:Parasite Load; D060888:Real-Time Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D013030:Spain / Q000453:epidemiology; D014171:Transients and Migrants; D014349:Trypanosoma cruzi / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification
chemical_list: D016054:DNA, Protozoan
keywords: Chagas disease; Discrete Typing Unit; Migration; Real-time PCR; Sequencing; Trypanosoma cruzi