Isolation and characterization of a new species of kobuvirus associated with cattle

J Gen Virol. 2003 Nov;84(Pt 11):3069-3077. doi: 10.1099/vir.0.19266-0.

Abstract

A cytopathic agent was isolated using Vero cells from the culture medium of HeLa cells that had been used for more than 30 years in our laboratory. This agent, termed U-1 strain, was serially passed in Vero cells with distinct CPE. Particles of U-1 strain negatively stained with phosphotungstic acid exhibited a distinct surface that resembled Aichi virus. The RNA genome of U-1 strain comprises 8374 nt, with a genome organization analogous to that of picornaviruses. Possible cleavage sites of the large ORF, which encoded a leader protein prior to the capsid protein region, were assigned following amino acid alignment with Aichi virus. The virus sequence had 33 and 75 % amino acid identity with the Aichi virus VP1 and 3D regions, respectively, but no more than 23 and 36 % with those of the prototype strains of other PICORNAVIRIDAE: The dendrogram based on the P1, P2 and P3 proteins indicated that U-1 strain is genetically included in the genus Kobuvirus but is distinct from Aichi virus. Of 72 cattle sera, 43 (59.7 %) were positive for neutralizing antibody against U-1 strain at a titre of 1 : 8 or more. However, sera from 190 humans, 242 monkeys, 139 pigs, 5 horses, 22 dogs and 9 cats did not neutralize U-1 strain at a 1 : 4 dilution. RNA corresponding to U-1 strain was detected in 12 (16.7 %) of 72 faecal samples from cattle by RT-PCR. These results indicated that U-1 strain, suspected to be a contaminant from calf sera, is a new species of the genus Kobuvirus, now termed bovine kobuvirus.

MeSH terms

  • 5' Untranslated Regions / chemistry
  • Animals
  • Antibodies, Viral / blood
  • Base Sequence
  • Cattle / virology*
  • Chlorocebus aethiops
  • Feces / virology
  • HeLa Cells
  • Humans
  • Molecular Sequence Data
  • Phylogeny
  • Picornaviridae / classification
  • Picornaviridae / immunology
  • Picornaviridae / isolation & purification*
  • RNA, Viral / analysis
  • Reverse Transcriptase Polymerase Chain Reaction
  • Vero Cells

Substances

  • 5' Untranslated Regions
  • Antibodies, Viral
  • RNA, Viral

Associated data

  • GENBANK/AB084788
  • GENBANK/AB097152
  • GENBANK/AB097153
  • GENBANK/AB097154
  • GENBANK/AB097155
  • GENBANK/AB097156
  • GENBANK/AB097157
  • GENBANK/AB097158
  • GENBANK/AB097159
  • GENBANK/AB097160
  • GENBANK/AB097161
  • GENBANK/AB097162
  • GENBANK/AB097163
  • GENBANK/AB097164
  • GENBANK/AB097165
  • GENBANK/AB097166