Resolving plasmid structures in Enterobacteriaceae using the MinION nanopore sequencer: assessment of MinION and MinION-Illumina hybrid data assembly approachesReport as inadecuate




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Reference: George, S, Pankhurst, L, Hubbard, A et al., (2017). Resolving plasmid structures in Enterobacteriaceae using the MinION nanopore sequencer: assessment of MinION and MinION/Illumina hybrid data assembly approaches. Microbial Genomics.Citable link to this page:

 

Resolving plasmid structures in Enterobacteriaceae using the MinION nanopore sequencer: assessment of MinION and MinION/Illumina hybrid data assembly approaches

Abstract: This study aimed to assess the feasibility of using the Oxford Nanopore Technologies (ONT) MinION long-read sequencer in reconstructing fully closed plasmid sequences from eight Enterobacteriaceae isolates of six different species with plasmid populations of varying complexity. Species represented were Escherichia coli, Klebsiella pneumoniae, Citrobacter freundii, Enterobacter cloacae, Serratia marcescens and Klebsiella oxytoca, with plasmid populations ranging from 1–11 plasmids with sizes of 2–330 kb. Isolates were sequenced using Illumina (short-read) and ONT’s MinION (long-read) platforms, and compared with fully resolved PacBio (long-read) sequence assemblies for the same isolates. We compared the performance of different assembly approaches including SPAdes, plasmidSPAdes, hybridSPAdes, Canu, Canu+Pilon (canuPilon) and npScarf in recovering the plasmid structures of these isolates by comparing with the gold-standard PacBio reference sequences. Overall, canuPilon provided consistently good quality assemblies both in terms of assembly statistics (N50, number of contigs) and assembly accuracy [presence of single nucleotide polymorphisms (SNPs)/indels with respect to the reference sequence]. For plasmid reconstruction, Canu recovered 70% of the plasmids in complete contigs, and combining three assembly approaches (Canu or canuPilon, hybridSPAdes and plasmidSPAdes) resulted in a total 78% recovery rate for all the plasmids. The analysis demonstrated the potential of using MinION sequencing technology to resolve important plasmid structures in Enterobacteriaceae species independent of and in conjunction with Illumina sequencing data. A consensus assembly derived from several assembly approaches could present significant benefit in accurately resolving the greatest number of plasmid structures.

Publication status:PublishedPeer Review status:Peer reviewedVersion:Publisher's versionDate of acceptance:2017-04-28 Funder: National Institute for Health Research   Funder: Public Health England   Funder: Wellcome Trust   Funder: UK Department of Health   Funder: Royal Society   Notes:© 2017 The Authors. This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/, which permits unrestricted use, distribution and reproduction in any medium, provided the original author and source are credited.

Bibliographic Details

Publisher: Microbiology Society

Publisher Website: http://www.microbiologyresearch.org/

Journal: Microbial Genomicssee more from them

Publication Website: http://mgen.microbiologyresearch.org/content/journal/mgen

Issue Date: 2017-06Identifiers

Doi: https://doi.org/10.1099/mgen.0.000118

Eissn: 2057-5858

Uuid: uuid:a2419a44-0d56-4dce-a2ba-27305b59829a

Urn: uri:a2419a44-0d56-4dce-a2ba-27305b59829a

Pubs-id: pubs:702996 Item Description

Type: journal-article;

Language: en

Version: Publisher's version

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Author: George, S - Oxford, MSD, NDM, NDM Experimental Medicine - - - Pankhurst, L - Oxford, MSD, NDM, NDM Experimental Medicine - - - Hu

Source: https://ora.ox.ac.uk/objects/uuid:a2419a44-0d56-4dce-a2ba-27305b59829a



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