A complete protocol for whole-genome sequencing of virus from clinical samples: Application to coronavirus OC43

Virology. 2019 May:531:141-148. doi: 10.1016/j.virol.2019.03.006. Epub 2019 Mar 9.

Abstract

Genome sequencing of virus has become a useful tool for better understanding of virus pathogenicity and epidemiological surveillance. Obtaining virus genome sequence directly from clinical samples is still a challenging task due to the low load of virus genetic material compared to the host DNA, and to the difficulty to get an accurate genome assembly. Here we introduce a complete sequencing and analyzing protocol called V-ASAP for Virus Amplicon Sequencing Assembly Pipeline. Our protocol is able to generate the viral dominant genome sequence starting from clinical samples. It is based on a multiplex PCR amplicon sequencing coupled with a reference-free analytical pipeline. This protocol was applied to 11 clinical samples infected with coronavirus OC43 (HcoV-OC43), and led to seven complete and two nearly complete genome assemblies. The protocol introduced here is shown to be robust, to produce a reliable sequence, and could be applied to other virus.

Keywords: Bioinformatics; Complete genome; Coronavirus; High-throughput sequencing.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Coronavirus Infections / virology*
  • Coronavirus OC43, Human / classification
  • Coronavirus OC43, Human / genetics*
  • Coronavirus OC43, Human / isolation & purification
  • Genome, Viral*
  • Humans
  • Multiplex Polymerase Chain Reaction
  • Whole Genome Sequencing / methods*