Culture-free genome-wide locus sequence typing (GLST) provides new perspectives on Trypanosoma cruzi dispersal and infection complexity

Simple item page
dc.careerEscuela de Ciencias Biológicases
dc.category.authorprincipales
dc.contributor.authorCostales Cordero, Jaime Alfredo
dc.contributor.authorOcaña Mayorga, Sofía Beatriz
dc.contributor.correspondingCostales Cordero, Jaime Alfredoes
dc.countryEcuadores
dc.date.accessioned2023-11-04T21:46:34Z
dc.date.available2023-11-04T21:46:34Z
dc.date.issued2020-12-16
dc.dedication.authorTCes
dc.description.abstractAnalysis of genetic polymorphism is a powerful tool for epidemiological surveillance and research. Powerful inference from pathogen genetic variation, however, is often restrained by limited access to representative target DNA, especially in the study of obligate parasitic species for which ex vivo culture is resource-intensive or bias-prone. Modern sequence capture methods enable pathogen genetic variation to be analyzed directly from host/vector material but are often too complex and expensive for resource-poor settings where infectious diseases prevail. This study proposes a simple, cost-effective ‘genome-wide locus sequence typing’ (GLST) tool based on massive parallel amplification of information hotspots throughout the target pathogen genome. The multiplexed polymerase chain reaction amplifies hundreds of different, user-defined genetic targets in a single reaction tube, and subsequent agarose gel-based clean-up and barcoding completes library preparation at under 4 USD per sample. Our study generates a flexible GLST primer panel design workflow for Trypanosoma cruzi, the parasitic agent of Chagas disease. We successfully apply our 203-target GLST panel to direct, culture-free metagenomic extracts from triatomine vectors containing a minimum of 3.69 pg/μl T. cruzi DNA and further elaborate on method performance by sequencing GLST libraries from T. cruzi reference clones representing discrete typing units (DTUs) TcI, TcIII, TcIV, TcV and TcVI. The 780 SNP sites we identify in the sample set repeatably distinguish parasites infecting sympatric vectors and detect correlations between genetic and geographic distances at regional (< 150 km) as well as continental scales. The markers also clearly separate TcI, TcIII, TcIV and TcV + TcVI and appear to distinguish multiclonal infections within TcI. We discuss the advantages, limitations and prospects of our method across a spectrum of epidemiological research.es
dc.facultyCiencias Exactas y Naturaleses
dc.id.author1710870252es
dc.id.author1711850931es
dc.id.type1es
dc.identifier.doihttps://doi.org/10.1371/journal.pgen.1009170es
dc.identifier.issn1553-7390
dc.identifier.urihttps://journals.plos.org/plosgenetics/article?id=10.1371/journal.pgen.1009170es
dc.identifier.urihttps://repositorio.puce.edu.ec/handle/123456789/6143
dc.indexed.databaseScimago Journal Rankes
dc.list.authorsSchwabl, P., Maiguashca, J., Costales, J., Ocaña-Mayorga, S., Segovia, M., Carrasco, H., et al.es
dc.magazine.pageRange1-10es
dc.magazine.titlePLoS Genetes
dc.magazine.volumeChapter16 (12)es
dc.rightsOpenAccesses
dc.statepublishedes
dc.subjectTécnicas genéticases
dc.subjectGenomases
dc.subjectProtozooes
dc.subjectTrypanosomaes
dc.titleCulture-free genome-wide locus sequence typing (GLST) provides new perspectives on Trypanosoma cruzi dispersal and infection complexityes
Files
License bundle
Now showing 1 - 1 of 1
No Thumbnail Available
Name:
license.txt
Size:
1.71 KB
Format:
Plain Text
Description:
Download
Collections
Artículos en revistas científicas indexadas (Journals)