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Artículo

Optimized multilocus sequence typing (MLST) scheme for Trypanosoma cruzi.

Diosque, PatricioIcon ; Tomasini, NicolásIcon ; Lauthier, Juan JoséIcon ; Messenger, Louisa Alexandra; Monje Rumi, Maria MercedesIcon ; Ragone, Paula GabrielaIcon ; Alberti D'Amato, Anahi Maiten; Perez Brandan, Cecilia MariaIcon ; Barnabe, Christian; Tibayrenc, Michel; Lewis, Michael David; Llewellyn, Martin Stephen; Miles, Michael Alexander; Yeo, Matthew
Fecha de publicación: 28/08/2014
Editorial: Public Library Of Science
Revista: Neglected Tropical Diseases
ISSN: 1935-2735
Idioma: Inglés
Tipo de recurso: Artículo publicado
Clasificación temática:
Enfermedades Infecciosas

Resumen

Trypanosoma cruzi, the aetiological agent of Chagas disease possess extensive genetic diversity. This has led to the development of a plethora of molecular typing methods for the identification of both the known major genetic lineages and for more fine scale characterization of different multilocus genotypes within these major lineages. Whole genome sequencing applied to large sample sizes is not currently viable and multilocus enzyme electrophoresis, the previous gold standard for T. cruzi typing, is laborious and time consuming. In the present work, we present an optimized Multilocus Sequence Typing (MLST) scheme, based on the combined analysis of two recently proposed MLST approaches. Here,thirteen concatenated gene fragments were applied to a panel of T. cruzi reference strains encompassing all known genetic lineages. Concatenation of 13 fragments allowed assignment of all strains to the predicted Discrete Typing Units (DTUs), or near-clades, with the exception of one strain that was an outlier for TcV, due to apparent loss of heterozygosity in one fragment. Monophyly for all DTUs, along with robust bootstrap support, was restored when this fragment was subsequently excluded from the analysis. All possible combinations of loci were assessed against predefined criteria with the objective of selecting the most appropriate combination of between two and twelve fragments, for an optimized MLST scheme. The optimum combination consisted of 7 loci and discriminated between all reference strains in the panel, with the majority supported by robust bootstrap values. Additionally, a reduced panel of just 4 gene fragments displayed high bootstrap values for DTU assignment and discriminated 21 out of 25 genotypes. We propose that the seven-fragment MLST scheme could be used as a gold standard for T. cruzi typing, against which other typing approaches, particularly single locus approaches or systematic PCR assays based on amplicon size, could be compared.
Palabras clave: Multilocus Sequence Typing , Trypanosoma Cruzi , Scheme of Optimization , Genetic Diversity
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info:eu-repo/semantics/openAccess Excepto donde se diga explícitamente, este item se publica bajo la siguiente descripción: Creative Commons Attribution 2.5 Unported (CC BY 2.5)
Identificadores
URI: http://hdl.handle.net/11336/7087
DOI: http://dx.doi.org/10.1371/journal.pntd.0003117
DOI: http://dx.doi.org/ 10.1371/journal.pntd.0003117
URL: http://journals.plos.org/plosntds/article?id=10.1371/journal.pntd.0003117
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Articulos(IPE)
Articulos de INST.DE PATOLOGIA EXPERIMENTAL
Citación
Diosque, Patricio; Tomasini, Nicolás; Lauthier, Juan José; Messenger, Louisa Alexandra; Monje Rumi, Maria Mercedes; et al.; Optimized multilocus sequence typing (MLST) scheme for Trypanosoma cruzi.; Public Library Of Science; Neglected Tropical Diseases; 8; 8; 28-8-2014; e3117
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