Isolation and genomic analyses of SARS-CoV-2 in Anhui Province, China.
Isolation and genomic analyses of SARS-CoV-2 in Anhui Province, China.
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DOI：
10.13242/j.cnki.bingduxuebao.003795
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发表时间：
2020-01-01
期刊：
影响因子：
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通讯作者：
He, J.
中科院分区：
文献类型：
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作者：
Li, Wei-wei;Sun, Yong;He, J.
SARS-CoV-2 作为 2019 年冠状病毒病（COVID-19）的病原体而受到全世界的关注。自2020年1月以来，中国安徽省报告确诊病例990例。我们希望探索 SARS-CoV-2 的分离和下一代测序 (NGS)。顺便，我们想了解中国安徽省SARS-CoV-2的变异情况，为疫苗的研发提供依据。 Vero 细胞被咽拭子样本感染。每天监测细胞的细胞病变效应（CPE）。通过逆转录聚合酶链反应（RT-PCR）检测细胞上清液中的 SARS-CoV-2 核酸。 SARS-CoV-2 的全基因组序列是通过基因组捕获和 NGS 生成的。序列比对由Lasergene MegAlign v7.1.0完成。系统发育树由PhyloSuite v1.2.1构建。该序列被上传到国家基因组数据中心以识别变异。 SARS-CoV-2分离显示在潜伏第四天出现CPE，并通过RT-PCR证实了该结果。该病毒基因组全长为 29, 860 bp，与其他 SARS-CoV-2 毒株具有超过 99% 的序列同一性，并有 9 个位置发生变化。蝙蝠中最相似的 SARS 样冠状病毒是云南 RaTG13 株（序列同一性为 96.8%）。这些数据表明，我们用 Vero 细胞分离了 SARS-CoV-2，并（首次）分析了中国安徽省 SARS-CoV-2 的全基因组序列。我们的结果将有助于了解 SARS-CoV-2 的特征。
SARS-CoV-2 was worldwide concerned as the pathogen of coronavirus disease 2019(COVID-19). Since January 2020, there were 990 confirmed cases reported in Anhui Province, China. We wish to explore the isolation and next-generation sequencing (NGS) of SARS-CoV-2. In the way, we wanted to understand the variation of SARS-CoV-2 in Anhui Province, China and provide a basis for the research and development of a vaccine. Vero cells were infected with throat swab samples. Cells were monitored daily for the cytopathic effect (CPE). Nucleic acids of SARS-CoV-2 in cell supernatants were detected by reverse transcription- polymerase chain reaction (RT-PCR). The whole genome sequence of SARS-CoV-2 was generated by genome capture and NGS. Sequence alignment was done by Lasergene MegAlign v7.1.0. Phylogenetic trees were constructed by PhyloSuite v1.2.1. The sequence was uploaded to the National Genomic Data Center to identify variations. SARS-CoV-2 isolation showed that a CPE appeared on the fourth day of incubation, and the result was confirmed by RT-PCR. The whole length of the virus genome was 29, 860 bp, which shared more than 99% sequence identity with other SARS-CoV-2 strains and varied in nine positions. The most similar SARS-like coronavirus in bats was the Yunnan RaTG13 strain (96.8% sequence identity). These data showed that we isolated SARS-CoV-2 with Vero cells and analyzed (for the first time) the whole genome sequence of SARS-CoV-2 in Anhui Province, China. Our results will aid understanding the characteristics of SARS-CoV-2.