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. 2007 Mar;124(1-2):78-87.
doi: 10.1016/j.virusres.2006.10.004. Epub 2006 Nov 7.

Isolation and sequence analysis of canine respiratory coronavirus

Kerstin Erles  1 , Kai-Biu Shiu, Joe Brownlie
Affiliations

Isolation and sequence analysis of canine respiratory coronavirus

Kerstin Erles et al. Virus Res. 2007 Mar.

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.

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Figures

Fig. 1
Fig. 1
Detection of CRCoV in HRT-18 cells by immunofluorescence assay using a polyclonal antiserum against bovine coronavirus.
Fig. 2
Fig. 2
Susceptibility of cell lines for infection with CRCoV. The cell lines were infected with CRCoV and total RNA was isolated 48 h post infection. RT-PCR for the detection of CRCoV nucleoprotein gene mRNA was performed using the primers BCV-LS and NP-6.
Fig. 3
Fig. 3
Comparison of the genomic organisation of the region located between the S and the E genes of BCoV strain Mebus and CRCoV strain 4182. Hatched boxes represent non-coding sequences. A–C indicate the three regions used for comparative nucleotide sequence analysis between CRCoV and BCoV strains.
Fig. 4
Fig. 4
Alignment of the predicted amino acid sequence of the putative CRCoV-4182 8.8 kDa protein and the putative CRCoV-G9142 4.9 kDa and 2.7 kDa proteins with the corresponding proteins of MHV-JHM and BCoV strains Mebus, LY-138 and BCO 43277.
Fig. 5
Fig. 5
Maximum parsimony phylogenetic tree of the non-structural region between the S and the E gene of CRCoV and other group 2 coronaviruses.
See this image and copyright information in PMC

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