Please use this identifier to cite or link to this item: http://www.repositorio.ufop.br/handle/123456789/3870
Title: Unequivocal identification of subpopulations in putative multiclonal Trypanosoma cruzi strains by FACs single cell sorting and genotyping.
Authors: Valadares, Helder Magno Silva
Pimenta, Juliana Ramos
Segatto, Marcela
Veloso, Vanja Maria
Gomes, Mônica Lúcia
Chiari, Egler
Gollob, Kenneth John
Bahia, Maria Terezinha
Lana, Marta de
Franco, Glória Regina
Machado, Carlos Renato
Pena, Sérgio Danilo Junho
Macedo, Andréa Mara
Issue Date: 2012
Citation: VALADARES, H. M. S. et al. Unequivocal identification of subpopulations in putative multiclonal Trypanosoma cruzi strains by FACs single cell sorting and genotyping. PLoS Neglected Tropical Diseases, v. 6, p. e1722-e1722, 2012. Disponível em: <http://www.plosntds.org/article/info%3Adoi%2F10.1371%2Fjournal.pntd.0001722>. Acesso em: 20 ago. 2014.
Abstract: Trypanosoma cruzi, the etiological agent of Chagas disease, is a polymorphic species. Evidence suggests that the majority of the T. cruzi populations isolated from afflicted humans, reservoir animals, or vectors are multiclonal. However, the extent and the complexity of multiclonality remain to be established, since aneuploidy cannot be excluded and current conventional cloning methods cannot identify all the representative clones in an infection. To answer this question, we adapted a methodology originally described for analyzing single spermatozoids, to isolate and study single T. cruzi parasites. Accordingly, the cloning apparatus of a Fluorescence-Activated Cell Sorter (FACS) was used to sort single T. cruzi cells directly into 96-wells microplates. Cells were then genotyped using two polymorphic genomic markers and four microsatellite loci. We validated this methodology by testing four T. cruzi populations: one control artificial mixture composed of two monoclonal populations – Silvio X10 cl1 (TcI) and Esmeraldo cl3 (TcII) – and three naturally occurring strains, one isolated from a vector (A316A R7) and two others derived from the first reported human case of Chagas disease. Using this innovative approach, we were able to successfully describe the whole complexity of these natural strains, revealing their multiclonal status. In addition, our results demonstrate that these T. cruzi populations are formed of more clones than originally expected. The method also permitted estimating of the proportion of each subpopulation of the tested strains. The single-cell genotyping approach allowed analysis of intrapopulation diversity at a level of detail not achieved previously, and may thus improve our comprehension of population structure and dynamics of T. cruzi. Finally, this methodology is capable to settle once and for all controversies on the issue of multiclonality.
URI: http://www.repositorio.ufop.br/handle/123456789/3870
metadata.dc.identifier.doi: https://doi.org/10.1371/journal.pntd.0001722
ISSN: 1935-2735
metadata.dc.rights.license: Nenhuma autorização é necessária para depósito dos artigos publicados pelos Periódicos PloS em repositório. Fonte: Plos <http://www.plos.org/open-access/> Acesso em: 14 nov. 2014.
Appears in Collections:DEFAR - Artigos publicados em periódicos

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