An amplicon-based sequencing framework for accurately measuring intrahost virus diversity using PrimalSeq and iVar

How viruses evolve within hosts can dictate infection outcomes; however, reconstructing this process is challenging. We evaluate our multiplexed amplicon approach, PrimalSeq, to demonstrate how virus concentration, sequencing coverage, primer mismatches, and replicates influence the accuracy of meas...

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Detalles Bibliográficos
Autores principales: Grubaugh, Nathan D., Gangavarapu, Karthik, Quick, Joshua, Matteson, Nathaniel L., De Jesus, Jaqueline Goes, Main, Bradley J., Tan, Amanda L., Paul, Lauren M., Brackney, Doug E., Grewal, Saran, Gurfield, Nikos, Van Rompay, Koen K. A., Isern, Sharon, Michael, Scott F., Coffey, Lark L., Loman, Nicholas J., Andersen, Kristian G.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2019
Materias:
Method
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6325816/
https://www.ncbi.nlm.nih.gov/pubmed/30621750
http://dx.doi.org/10.1186/s13059-018-1618-7
Descripción
Sumario:How viruses evolve within hosts can dictate infection outcomes; however, reconstructing this process is challenging. We evaluate our multiplexed amplicon approach, PrimalSeq, to demonstrate how virus concentration, sequencing coverage, primer mismatches, and replicates influence the accuracy of measuring intrahost virus diversity. We develop an experimental protocol and computational tool, iVar, for using PrimalSeq to measure virus diversity using Illumina and compare the results to Oxford Nanopore sequencing. We demonstrate the utility of PrimalSeq by measuring Zika and West Nile virus diversity from varied sample types and show that the accumulation of genetic diversity is influenced by experimental and biological systems. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13059-018-1618-7) contains supplementary material, which is available to authorized users.

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