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

Abstract:

: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.

journal_name

Genome Biol

journal_title

Genome biology

authors

Grubaugh ND,Gangavarapu K,Quick J,Matteson NL,De Jesus JG,Main BJ,Tan AL,Paul LM,Brackney DE,Grewal S,Gurfield N,Van Rompay KKA,Isern S,Michael SF,Coffey LL,Loman NJ,Andersen KG

doi

10.1186/s13059-018-1618-7

subject

Has Abstract

pub_date

2019-01-08 00:00:00

pages

8

issue

1

eissn

1474-7596

issn

1474-760X

pii

10.1186/s13059-018-1618-7

journal_volume

20

pub_type

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