Unequivocal identification of subpopulations in putative multiclonal Trypanosoma cruzi strains by FACs single cell sorting and genotyping.

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dc.contributor.authorValadares, Helder Magno Silva
dc.contributor.authorPimenta, Juliana Ramos
dc.contributor.authorSegatto, Marcela
dc.contributor.authorVeloso, Vanja Maria
dc.contributor.authorGomes, Mônica Lúcia
dc.contributor.authorChiari, Egler
dc.contributor.authorGollob, Kenneth John
dc.contributor.authorBahia, Maria Terezinha
dc.contributor.authorLana, Marta de
dc.contributor.authorFranco, Glória Regina
dc.contributor.authorMachado, Carlos Renato
dc.contributor.authorPena, Sérgio Danilo Junho
dc.contributor.authorMacedo, Andréa Mara
dc.date.accessioned2014-11-14T19:36:45Z
dc.date.available2014-11-14T19:36:45Z
dc.date.issued2012
dc.description.abstractTrypanosoma 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.pt_BR
dc.identifier.citationVALADARES, 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.pt_BR
dc.identifier.doihttps://doi.org/10.1371/journal.pntd.0001722
dc.identifier.issn1935-2735
dc.identifier.urihttp://www.repositorio.ufop.br/handle/123456789/3870
dc.language.isoen_USpt_BR
dc.rights.licenseNenhuma 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.pt_BR
dc.titleUnequivocal identification of subpopulations in putative multiclonal Trypanosoma cruzi strains by FACs single cell sorting and genotyping.pt_BR
dc.typeArtigo publicado em periodicopt_BR
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