Genomic analysis of canine pneumoviruses and canine respiratory coronavirus from New Zealand.

IF 16.4 1区 化学 Q1 CHEMISTRY, MULTIDISCIPLINARY
M. Dunowska, G. More, PJ Biggs, N. Cave
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引用次数: 0

Abstract

AIMS To isolate canine respiratory coronavirus (CRCoV) and canine pneumovirus (CnPnV) in cell culture and to compare partial genomic sequences of CRCoV and CnPnV from New Zealand with those from other countries. METHODS Oropharyngeal swab samples from dogs affected by canine infectious respiratory disease syndrome that were positive for CnPnV (n = 15) or CRCoV (n = 1) by virus-specific reverse transcriptase quantitative PCR (RT-qPCR) in a previous study comprised the starting material. Virus isolation was performed in HRT-18 cells for CRCoV and RAW 264.7 and Vero cells for CnPnV. The entire sequence of CnPnV G protein (1,266 nucleotides) and most (8,063/9,707 nucleotides) of the 3' region of CRCoV that codes for 10 structural and accessory proteins were amplified and sequenced. The sequences were analysed and compared with other sequences available in GenBank using standard molecular tools including phylogenetic analysis. RESULTS Virus isolation was unsuccessful for both CRCoV and CnPnV. Pneumovirus G protein was amplified from 3/15 (20%) samples that were positive for CnPnV RNA by RT-qPCR. Two of these (NZ-048 and NZ-049) were 100% identical to each other, and 90.9% identical to the third one (NZ-007). Based on phylogenetic analysis of the G protein gene, CnPnV NZ-048 and NZ-049 clustered with sequences from the USA, Thailand and Italy in group A, and CnPnV NZ-007 clustered with sequences from the USA in group B. The characteristics of the predicted genes (length, position) and their putative protein products (size, predicted structure, presence of N- and O-glycosylation sites) of the New Zealand CRCoV sequence were consistent with those reported previously, except for the region located between open reading frame (ORF)3 (coding for S protein) and ORF6 (coding for E protein). The New Zealand virus was predicted to encode 5.9 kDa, 27 kDa and 12.7 kDa proteins, which differed from the putative coding capacity of this region reported for CRCoV from other countries. CONCLUSIONS This report represents the first characterisation of partial genomic sequences of CRCoV and CnPnV from New Zealand. Our results suggest that the population of CnPnV circulating in New Zealand is not homogeneous, and that the viruses from two clades described overseas are also present here. Limited conclusions can be made based on only one CRCoV sequence, but the putative differences in the coding capacity of New Zealand CRCoV support the previously reported variability of this region. The reasons for such variability and its biological implications need to be further elucidated.
新西兰犬肺炎病毒和犬呼吸道冠状病毒的基因组分析。
目的在细胞培养中分离犬呼吸道冠状病毒(CRCoV)和犬肺炎病毒(CnPnV),并比较新西兰与其他国家的 CRCoV 和 CnPnV 的部分基因组序列。方法以犬传染性呼吸道疾病综合征患犬的口咽拭子样本为起始材料,这些患犬在之前的研究中通过病毒特异性逆转录酶定量 PCR(RT-qPCR)检测出 CnPnV(n = 15)或 CRCoV(n = 1)呈阳性。CRCoV 的病毒分离在 HRT-18 细胞中进行,CnPnV 的病毒分离在 RAW 264.7 和 Vero 细胞中进行。对 CnPnV G 蛋白的整个序列(1,266 个核苷酸)和 CRCoV 编码 10 个结构蛋白和附属蛋白的 3' 区域的大部分序列(8,063/9,707 个核苷酸)进行了扩增和测序。结果CRCoV和CnPnV的病毒分离均未成功。通过 RT-qPCR,从 3/15 份(20%)对 CnPnV RNA 呈阳性的样本中扩增出了肺炎病毒 G 蛋白。其中两个样本(NZ-048 和 NZ-049)100%相同,第三个样本(NZ-007)90.9%相同。根据 G 蛋白基因的系统进化分析,CnPnV NZ-048 和 NZ-049 与来自美国、泰国和意大利的序列聚为 A 组,CnPnV NZ-007 与来自美国的序列聚为 B 组。新西兰 CRCoV 序列中预测基因的特征(长度、位置)及其假定蛋白产物的特征(大小、预测结构、N-和 O-糖基化位点的存在)与之前报道的一致,但位于开放阅读框(ORF)3(编码 S 蛋白)和 ORF6(编码 E 蛋白)之间的区域除外。据预测,新西兰病毒可编码 5.9 kDa、27 kDa 和 12.7 kDa 蛋白,这与其他国家报告的 CRCoV 在该区域的推测编码能力不同。我们的研究结果表明,在新西兰流行的 CnPnV 群体并不均匀,国外描述的两个支系的病毒在这里也有存在。仅根据一个 CRCoV 序列得出的结论有限,但新西兰 CRCoV 编码能力的推测差异支持了之前报告的该地区的变异性。这种变异的原因及其生物学意义有待进一步阐明。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Accounts of Chemical Research
Accounts of Chemical Research 化学-化学综合
CiteScore
31.40
自引率
1.10%
发文量
312
审稿时长
2 months
期刊介绍: Accounts of Chemical Research presents short, concise and critical articles offering easy-to-read overviews of basic research and applications in all areas of chemistry and biochemistry. These short reviews focus on research from the author’s own laboratory and are designed to teach the reader about a research project. In addition, Accounts of Chemical Research publishes commentaries that give an informed opinion on a current research problem. Special Issues online are devoted to a single topic of unusual activity and significance. Accounts of Chemical Research replaces the traditional article abstract with an article "Conspectus." These entries synopsize the research affording the reader a closer look at the content and significance of an article. Through this provision of a more detailed description of the article contents, the Conspectus enhances the article's discoverability by search engines and the exposure for the research.
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