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Parasitol Int. 2017 Apr;66(2):83-88. doi: 10.1016/j.parint.2016.12.003. Epub 2016 Dec 7.

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

Author information

1
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.
2
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.
3
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.
4
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.
5
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.
6
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.
7
ISGlobal, Barcelona Centre for International Health Research (CRESIB), Roselló 134-4°, 08036 Barcelona, Spain.
8
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.
9
Secció de Parasitologia, Departament de Biologia, Sanitat i Medi Ambient, Facultat de Farmàcia, Universitat de Barcelona, Av. Joan XXIII 27-31, 08028 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.

KEYWORDS:

Chagas disease; Discrete Typing Unit; Migration; Real-time PCR; Sequencing; Trypanosoma cruzi

PMID:
27940065
DOI:
10.1016/j.parint.2016.12.003
[Indexed for MEDLINE]

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