Identification and genetic characterization of bovine enterovirus by combination of two next generation sequencing platforms

J Virol Methods. 2018 Oct:260:21-25. doi: 10.1016/j.jviromet.2018.07.002. Epub 2018 Jul 4.

Abstract

Prompt and accurate diagnosis is warranted for infectious diseases of domestic animals which may have a significant impact on animal production or clinical practice. In this study, the identification and genetic characterization of a bovine enterovirus (BEV) strain isolated from a calf with diarrhea, are described. Two different next generation sequencing platforms were employed. Shotgun metagenomic accomplished by MinION sequencing (Oxford Nanopore Technologies) allowed the identification of BEV RNA from a cell-culture isolate. BEV was then confirmed by a specific real time RT-PCR assay. To achieve the whole genome of this isolate, sequence reads obtained by MinION were coupled with those originating from NextSeq500 (Illumina). Genomic relatedness and phylogeny with extant BEV strains is also reported. Overall, this manuscript highlights the use of the portable MinION sequence technology as a tool for support diagnostics in veterinary practice.

Keywords: Illumina; MinION; NGS; bovine; characterization; diagnosis; enterovirus.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cattle
  • Chlorocebus aethiops
  • Diarrhea / diagnosis*
  • Diarrhea / veterinary
  • Enterovirus Infections / diagnosis*
  • Enterovirus Infections / veterinary
  • Enterovirus, Bovine / genetics*
  • Enterovirus, Bovine / isolation & purification*
  • Feces / virology
  • High-Throughput Nucleotide Sequencing*
  • Phylogeny
  • RNA, Viral / isolation & purification
  • Sequence Analysis, RNA
  • Vero Cells
  • Whole Genome Sequencing

Substances

  • RNA, Viral