NHC Key Laboratory of Biosafety, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, China.
Key Laboratory of Etiology and Epidemiology of Emerging Infectious Diseases in Universities of Shandong, Shandong First Medical University and Shandong Academy of Medical Sciences, Tai'an, China.
Lancet. 2020 Feb 22;395(10224):565-574. doi: 10.1016/S0140-6736(20)30251-8. Epub 2020 Jan 30.
BACKGROUND: In late December, 2019, patients presenting with viral pneumonia due to an unidentified microbial agent were reported in Wuhan, China. A novel coronavirus was subsequently identified as the causative pathogen, provisionally named 2019 novel coronavirus (2019-nCoV). As of Jan 26, 2020, more than 2000 cases of 2019-nCoV infection have been confirmed, most of which involved people living in or visiting Wuhan, and human-to-human transmission has been confirmed. METHODS: We did next-generation sequencing of samples from bronchoalveolar lavage fluid and cultured isolates from nine inpatients, eight of whom had visited the Huanan seafood market in Wuhan. Complete and partial 2019-nCoV genome sequences were obtained from these individuals. Viral contigs were connected using Sanger sequencing to obtain the full-length genomes, with the terminal regions determined by rapid amplification of cDNA ends. Phylogenetic analysis of these 2019-nCoV genomes and those of other coronaviruses was used to determine the evolutionary history of the virus and help infer its likely origin. Homology modelling was done to explore the likely receptor-binding properties of the virus. FINDINGS: The ten genome sequences of 2019-nCoV obtained from the nine patients were extremely similar, exhibiting more than 99·98% sequence identity. Notably, 2019-nCoV was closely related (with 88% identity) to two bat-derived severe acute respiratory syndrome (SARS)-like coronaviruses, bat-SL-CoVZC45 and bat-SL-CoVZXC21, collected in 2018 in Zhoushan, eastern China, but were more distant from SARS-CoV (about 79%) and MERS-CoV (about 50%). Phylogenetic analysis revealed that 2019-nCoV fell within the subgenus Sarbecovirus of the genus Betacoronavirus, with a relatively long branch length to its closest relatives bat-SL-CoVZC45 and bat-SL-CoVZXC21, and was genetically distinct from SARS-CoV. Notably, homology modelling revealed that 2019-nCoV had a similar receptor-binding domain structure to that of SARS-CoV, despite amino acid variation at some key residues. INTERPRETATION: 2019-nCoV is sufficiently divergent from SARS-CoV to be considered a new human-infecting betacoronavirus. Although our phylogenetic analysis suggests that bats might be the original host of this virus, an animal sold at the seafood market in Wuhan might represent an intermediate host facilitating the emergence of the virus in humans. Importantly, structural analysis suggests that 2019-nCoV might be able to bind to the angiotensin-converting enzyme 2 receptor in humans. The future evolution, adaptation, and spread of this virus warrant urgent investigation. FUNDING: National Key Research and Development Program of China, National Major Project for Control and Prevention of Infectious Disease in China, Chinese Academy of Sciences, Shandong First Medical University.
背景:2019 年 12 月,中国武汉报告了多例不明原因的病毒性肺炎病例。随后鉴定出一种新型冠状病毒,暂命名为 2019 新型冠状病毒(2019-nCoV)。截至 2020 年 1 月 26 日,已确认超过 2000 例 2019-nCoV 感染病例,其中大多数涉及居住或访问武汉的人,并且已确认人际传播。
方法:我们对 9 名住院患者的支气管肺泡灌洗液样本和培养物进行了下一代测序,其中 8 名患者曾前往武汉华南海鲜市场。从这些个体中获得了完整和部分 2019-nCoV 基因组序列。通过桑格测序连接病毒基因簇以获得全长基因组,通过快速扩增 cDNA 末端确定末端区域。对这些 2019-nCoV 基因组和其他冠状病毒的系统发育分析用于确定病毒的进化历史,并有助于推断其可能的起源。同源建模用于探索病毒可能的受体结合特性。
结果:从 9 名患者中获得的 2019-nCoV 的 10 个基因组序列非常相似,显示出超过 99.98%的序列同一性。值得注意的是,2019-nCoV 与 2018 年在中国东部舟山采集的两种蝙蝠源性严重急性呼吸系统综合征(SARS)样冠状病毒蝙蝠-SL-CoVZC45 和蝙蝠-SL-CoVZXC21 密切相关(同源性为 88%),但与 SARS-CoV(约 79%)和 MERS-CoV(约 50%)的关系较远。系统发育分析显示,2019-nCoV 属于β冠状病毒属的Sarbecovirus 亚属,与最接近的亲缘病毒蝙蝠-SL-CoVZC45 和蝙蝠-SL-CoVZXC21 相比,其分支长度较长,与 SARS-CoV 相比,遗传上也有所不同。值得注意的是,尽管在一些关键残基上存在氨基酸变异,但同源建模显示 2019-nCoV 具有与 SARS-CoV 相似的受体结合域结构。
结论:2019-nCoV 与 SARS-CoV 的差异足以被视为一种新的感染人类的β冠状病毒。尽管我们的系统发育分析表明蝙蝠可能是该病毒的原始宿主,但在武汉海鲜市场销售的一种动物可能代表一种中间宿主,有助于该病毒在人类中的出现。重要的是,结构分析表明 2019-nCoV 可能能够与人类的血管紧张素转换酶 2 受体结合。该病毒的未来进化、适应和传播值得紧急调查。
资金:国家重点研发计划、国家重大传染病防治科技重大专项、中国科学院、山东第一医科大学。
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