Isolation and sequence analysis of canine respiratory coronavirus

Abstract

Canine respiratory coronavirus (CRCoV) has frequently been detected in respiratory samples from dogs by RT-PCR. In this report the first successful isolation of CRCoV from a dog with respiratory disease is described. The isolate CRCoV-4182 was cultured in HRT-18 cells but failed to replicate in a number of other cell lines. The nucleotide sequence of the 3'-terminal portion of the CRCoV genome was determined including all open reading frames from the NS2 gene to the N gene. Comparison with other coronavirus sequences showed a high similarity to bovine coronavirus (BCoV). The region between the spike and the E gene was found to be the most variable and was used for phylogenetic analysis of several CRCoV strains. CRCoV-4182 showed a mutation within the non-structural protein region downstream of the S gene leading to the translation of an 8.8 kDa putative protein comprising a fusion of the equivalent of the BCoV 4.9 kDa protein to a truncated version of the BCoV 4.8 kDa protein. The culture of CRCoV will enable analysis of the expression and function of this and other CRCoV proteins as well as allowing the study of the role of CRCoV in the aetiology of canine infectious respiratory disease.

Evolutionary dynamics of bovine coronaviruses: natural selection pattern of the spike gene implies adaptive evolution of the strains

“…BCoV, together with human coronavirus OC43 (HCoV-OC43), equine coronavirus (ECoV) and porcine haemagglutinating encephalomyelitis virus (PHEV) belong to the virus species Betacoronavirus 1 of the lineage A of the genus Betacoronavirus (de Groot et al, 2012). A recently isolated canine respiratory coronavirus (CRCoV) has also shown a high genetic similarity to Betacoronavirus 1 (Erles et al, 2007).…”

Epidemic of Equine Coronavirus at Obihiro Racecourse, Hokkaido, Japan in 2012

“…The p4.7 to p12.7 genes in ECoV, which corresponds to p4.9, p4.8 and p12.7 genes in BCoV [7], are between S and envelope genes in Betacoronavirus 1. In this region, CRCoVs have 3 (p4.9, p2.7 and p12.8) genes or 2 (p8.8 and p12.8) genes depending on virus strains [6,13]. The functions of these non-structural proteins are unclear in Betacoronairus 1.…”

Simultaneous Detection of Canine Respiratory Disease Associated Viruses by a Multiplex Reverse Transcription-Polymerase Chain Reaction Assay

“…Each clinical sample was tested by RT-PCR specific for each of the 3 target viruses one at a time. The primers used in RT-PCR for the detection of the CDV nucleoprotein (NP) and the CRCoV NP were PP-I and BCV-LS/NP6, respectively [9,10]. The product sizes were 287 bp and 500 bp, respectively.…”