Erles Kerstin, Shiu Kai-Biu, Brownlie Joe
The Royal Veterinary College, Department of Pathology and Infectious Diseases, Hawkshead Lane, Hatfield AL9 7TA, United Kingdom.
Virus Res. 2007 Mar;124(1-2):78-87. doi: 10.1016/j.virusres.2006.10.004. Epub 2006 Nov 7.
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.
通过逆转录聚合酶链反应（RT-PCR），犬呼吸道冠状病毒（CRCoV）经常在犬类的呼吸道样本中被检测到。在本报告中，描述了首次从一只患有呼吸道疾病的犬成功分离出CRCoV。分离株CRCoV-4182在HRT-18细胞中培养，但未能在许多其他细胞系中复制。测定了CRCoV基因组3'末端部分的核苷酸序列，包括从NS2基因到N基因的所有开放阅读框。与其他冠状病毒序列的比较显示与牛冠状病毒（BCoV）高度相似。发现刺突蛋白和E基因之间的区域是最可变的，并用于几种CRCoV毒株的系统发育分析。CRCoV-4182在S基因下游的非结构蛋白区域内显示出一个突变，导致翻译出一种8.8 kDa的推定蛋白，该蛋白由相当于BCoV 4.9 kDa蛋白与BCoV 4.8 kDa蛋白的截短版本融合而成。CRCoV的培养将有助于分析该蛋白和其他CRCoV蛋白的表达和功能，以及研究CRCoV在犬传染性呼吸道疾病病因学中的作用。
