Novel Canine Coronavirus Isolated from a Hospitalized Patient With Pneumonia in East Malaysia
Novel Canine Coronavirus Isolated from a Hospitalized Patient With Pneumonia in East Malaysia
复制标题
DOI：
10.1093/cid/ciab456
复制
发表时间：
2021-07-14
影响因子：
11.8
通讯作者：
Gray, Gregory C.
中科院分区：
文献类型：
--
作者：
Vlasova, Anastasia N.;Diaz, Annika;Gray, Gregory C.
这是从肺炎患儿分离的新型犬-猫重组冠状病毒的首次完整基因组表征。与严重急性呼吸综合征冠状病毒相似，这种新型病毒具有一些独特的遗传特征，提示最近的人畜共患传播。背景在验证一种高度敏感的泛种冠状病毒（CoV）套式逆转录聚合酶链反应（RT-PCR）检测方法时，我们在2017-2018年马来西亚沙捞越301例肺炎住院患者中的8例（2.5%）的鼻咽拭子样本中发现了犬冠状病毒（CCoV）RNA。大多数患者是生活在农村地区的儿童，经常接触驯养动物和野生动物。方法采用通用型和种特异性的CoV和CCoV一步法RT-PCR检测方法，并在A72犬细胞中进行病毒分离。使用桑格方法进行全基因组测序。结果8份标本中有2份经不太敏感的一步法RT-PCR检测证实含有足够量的CCoV，1份标本在A72细胞中显示细胞病变。引起细胞病变效应的病毒的全基因组测序将其鉴定为一种新型犬-猫重组冠状病毒（基因型II），我们将其命名为CCoV-人肺炎（HuPn）-2018。CCoV-HuPn-2018基因组的大部分与CCoV TN-449更密切相关，而其S基因与CCoV-UCD-1（S1结构域）和猫CoV WSU 79-1683（S2结构域）具有显著更高的序列同一性。CCoV-HuPn-2018的独特之处在于N蛋白中的36个核苷酸（12个氨基酸）缺失以及全长和截短的7 b非结构蛋白的存在，这可能具有临床意义。结论这是首次从人类肺炎患者中分离到一株新型犬-猫重组冠状病毒。如果确认为病原体，它可能代表已知引起人类疾病的第八种独特冠状病毒。我们的研究结果强调了动物冠状病毒对公共卫生的威胁，以及对它们进行更好监测的必要性。
This is the first complete genome characterization of a novel canine-feline recombinant alphacoronavirus isolated from a child with pneumonia. Similar to severe acute respiratory syndrome coronavirus, this novel virus possesses some unique genetic features suggestive of recent zoonotic transmission.Background During the validation of a highly sensitive panspecies coronavirus (CoV) seminested reverse-transcription polymerase chain reaction (RT-PCR) assay, we found canine CoV (CCoV) RNA in nasopharyngeal swab samples from 8 of 301 patients (2.5%) hospitalized with pneumonia during 2017-2018 in Sarawak, Malaysia. Most patients were children living in rural areas with frequent exposure to domesticated animals and wildlife. Methods Specimens were further studied with universal and species-specific CoV and CCoV 1-step RT-PCR assays, and viral isolation was performed in A72 canine cells. Complete genome sequencing was conducted using the Sanger method. Results Two of 8 specimens contained sufficient amounts of CCoVs as confirmed by less-sensitive single-step RT-PCR assays, and 1 specimen demonstrated cytopathic effects in A72 cells. Complete genome sequencing of the virus causing cytopathic effects identified it as a novel canine-feline recombinant alphacoronavirus (genotype II) that we named CCoV-human pneumonia (HuPn)-2018. Most of the CCoV-HuPn-2018 genome is more closely related to a CCoV TN-449, while its S gene shared significantly higher sequence identity with CCoV-UCD-1 (S1 domain) and a feline CoV WSU 79-1683 (S2 domain). CCoV-HuPn-2018 is unique for a 36-nucleotide (12-amino acid) deletion in the N protein and the presence of full-length and truncated 7b nonstructural protein, which may have clinical relevance. Conclusions This is the first report of a novel canine-feline recombinant alphacoronavirus isolated from a human patient with pneumonia. If confirmed as a pathogen, it may represent the eighth unique coronavirus known to cause disease in humans. Our findings underscore the public health threat of animal CoVs and a need to conduct better surveillance for them.