Isolation and characterization of a new species of kobuvirus associated with cattle.
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.
Department of Microbiology, Aichi Prefectural Institute of Public Health, 7-6 Nagare, Tsujimachi, Kita-ku, Nagoya, Aichi 4628576, Japan.
Teruo
T
Yamashita
Department of Microbiology, Aichi Prefectural Institute of Public Health, 7-6 Nagare, Tsujimachi, Kita-ku, Nagoya, Aichi 4628576, Japan.
Miyabi
M
Ito
Department of Microbiology, Aichi Prefectural Institute of Public Health, 7-6 Nagare, Tsujimachi, Kita-ku, Nagoya, Aichi 4628576, Japan.
Yuka
Y
Kabashima
Department of Microbiology, Aichi Prefectural Institute of Public Health, 7-6 Nagare, Tsujimachi, Kita-ku, Nagoya, Aichi 4628576, Japan.
Hideaki
H
Tsuzuki
Department of Microbiology, Aichi Prefectural Institute of Public Health, 7-6 Nagare, Tsujimachi, Kita-ku, Nagoya, Aichi 4628576, Japan.
Akira
A
Fujiura
Department of Microbiology, Aichi Prefectural Institute of Public Health, 7-6 Nagare, Tsujimachi, Kita-ku, Nagoya, Aichi 4628576, Japan.
Kenji
K
Sakae
D020121:5' Untranslated Regions / Q000737:chemistry; D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D001483:Base Sequence; D002417:Cattle / Q000821:virology*; D002522:Chlorocebus aethiops; D005243:Feces / Q000821:virology; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D010802:Phylogeny; D010849:Picornaviridae / Q000145:classification / Q000276:immunology / Q000302:isolation & purification*; D012367:RNA, Viral / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014709:Vero Cells
D020121:5' Untranslated Regions; D000914:Antibodies, Viral; D012367:RNA, Viral