Whole genome sequencing of Enterovirus species C isolates by high-throughput sequencing: Development of generic primers

Public Deposited
Analytics

Downloadable Content

Download PDF
Request Version for Screen Reader
Creator
  • Bessaud, M.
    • Other Affiliation: Institut Pasteur
  • Sadeuh-Mba, S.A.
    • Other Affiliation: Centre Pasteur du Cameroun
  • Joffret, M.-L.
    • Other Affiliation: Institut Pasteur
  • Razafindratsimandresy, R.
    • Other Affiliation: Institut Pasteur de Madagascar
  • Polston, P.
    • Affiliation: Gillings School of Global Public Health, Department of Health Behavior
    • Other Affiliation: Institut Pasteur
  • Volle, R.
    • Other Affiliation: Institut Pasteur
  • Rakoto-Andrianarivelo, M.
    • Other Affiliation: Université Ankatso
  • Blondel, B.
    • Other Affiliation: Institut Pasteur
  • Njouom, R.
    • Other Affiliation: Centre Pasteur du Cameroun
  • Delpeyroux, F.
    • Other Affiliation: Institut Pasteur
Abstract
  • Enteroviruses are among the most common viruses infecting humans and can cause diverse clinical syndromes ranging from minor febrile illness to severe and potentially fatal diseases. Enterovirus species C (EV-C) consists of more than 20 types, among which the three serotypes of polioviruses, the etiological agents of poliomyelitis, are included. Biodiversity and evolution of EV-C genomes are shaped by frequent recombination events. Therefore, identification and characterization of circulating EV-C strains require the sequencing of different genomic regions. A simple method was developed to quickly sequence the entire genome of EV-C isolates. Four overlapping fragments were produced separately by RT-PCR performed with generic primers. The four amplicons were then pooled and purified prior to being sequenced by a high-throughput technique. The method was assessed on a panel of EV-Cs belonging to a wide-range of types. It can be used to determine full-length genome sequences through de novo assembly of thousands of reads. It was also able to discriminate reads from closely related viruses in mixtures. By decreasing the workload compared to classical Sanger-based techniques, this method will serve as a precious tool for sequencing large panels of EV-Cs isolated in cell cultures during environmental surveillance or from patients, including vaccine-derived polioviruses.
Date of publication
Keyword
DOI
Resource type
  • Article
Rights statement
  • In Copyright
License
  • Attribution 3.0 United States
Journal title
  • Frontiers in Microbiology
Journal volume
  • 7
Journal issue
  • AUG
Language
  • English
Version
  • Publisher
ISSN
  • 1664-302X
Publisher
  • Frontiers Media S.A.

Relations

Parents:

This work has no parents.

In Collection:

Items

Thumbnail Title Date Uploaded Visibility Actions
file details fmicb-07-01294.pdf 2021-06-21 Public Download