Full-length genome sequences of porcine epidemic diarrhoea virus strain CV777; Use of NGS to analyse genomic and sub-genomic RNAs

PLoS One. 2018 Mar 1;13(3):e0193682. doi: 10.1371/journal.pone.0193682. eCollection 2018.

Abstract

Porcine epidemic diarrhoea virus, strain CV777, was initially characterized in 1978 as the causative agent of a disease first identified in the UK in 1971. This coronavirus has been widely distributed among laboratories and has been passaged both within pigs and in cell culture. To determine the variability between different stocks of the PEDV strain CV777, sequencing of the full-length genome (ca. 28kb) has been performed in 6 different laboratories, using different protocols. Not surprisingly, each of the different full genome sequences were distinct from each other and from the reference sequence (Accession number AF353511) but they are >99% identical. Unique and shared differences between sequences were identified. The coding region for the surface-exposed spike protein showed the highest proportion of variability including both point mutations and small deletions. The predicted expression of the ORF3 gene product was more dramatically affected in three different variants of this virus through either loss of the initiation codon or gain of a premature termination codon. The genome of one isolate had a substantially rearranged 5´-terminal sequence. This rearrangement was validated through the analysis of sub-genomic mRNAs from infected cells. It is clearly important to know the features of the specific sample of CV777 being used for experimental studies.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Coronavirus Infections / virology*
  • Evolution, Molecular
  • Genome, Viral
  • High-Throughput Nucleotide Sequencing / methods*
  • Open Reading Frames
  • Phylogeny
  • Point Mutation
  • Porcine epidemic diarrhea virus / genetics*
  • Porcine epidemic diarrhea virus / isolation & purification
  • RNA, Viral / chemistry
  • RNA, Viral / genetics
  • Sequence Analysis, RNA / methods*
  • Sequence Deletion
  • Swine
  • Swine Diseases / virology*

Substances

  • RNA, Viral

Grants and funding

Financial support was received from the CoVet Lab (a partnership of national veterinary public health institutes from Denmark, France, The Netherlands, Sweden and the United Kingdom) to AB, WvdP, FS and YB. The study has also been funded, in part, by the EU Horizon 2020 program (European Commission Grant Agreement No. 643476 “COMPARE”) for SB and YB. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.