广东省首例新型冠状病毒的分离与鉴定  

First Isolation and Identification of SARS-CoV-2 in Guangdong Province,China

作　　者：张欢[1] 郑焕英[1] 邹丽容[1] 刘哲[2] 梁丽君[1] 彭晓放 张薇[1] 柯昌文[1] 武婕[1] ZHANG Huan;ZHENG Huanying;ZOU Lirong;LIU Zhe;LIANG Lijun;PENG Xiaofang;ZHANG Wei;KE Changwen;WU Jie(Guangdong Provincial Center for Disease Control and Prevention,Guangzhou 511430,China;Institute of Public Health,Guangdong Provincial Center for Disease Control and Prevention,Guangzhou 511430,China)

机构地区：[1]广东省疾病预防控制中心,广州511430 [2]广东省疾病预防控制中心,广东省公共卫生研究院,广州511430

出　　处：《病毒学报》2020年第2期155-159,共5页Chinese Journal of Virology

基　　金：广东省自然科学基金-面上项目(项目号:2019A1515012121);题目:轨道交通系统中流感病毒的周期变化规律研究。

摘　　要：2020年1月,广东省陆续从湖北省武汉市输入多例疑似新冠状病毒(SARS-CoV-2)感染的病例,经检测为SARS-CoV-2核酸阳性的实验室确诊病例。为进一步了解SARS-CoV-2毒力,满足药物研发和疫情防控等需求,并建立SARS-CoV-2分离方法为后续研究打下基础,本研究使用Vero-E6细胞,选择一例经Real-time RT-PCR鉴定为SARS-CoV-2阳性的肺泡灌洗液进行接种,逐日观察,肺泡灌洗液接种的细胞管4d出现疑似细胞病变(Cytopathic effect,CPE),再进行第二代接种2d出现病变,提取核酸进行鉴定,为SARS-CoV-2阳性。经二代测序成功获得全基因组序列,进化分析结果显示本次分离的毒株为SARS-CoV-2。In January 2020,Guangdong Province,China imported several suspected cases with SARS-CoV-2 from Wuhan City,Hubei Province,China,which were detected as SARS-CoV-2 positive in laboratory. To further understand the SARS-CoV-2 virulence,as well as drug development and epidemic prevention and control needs,we established a SARS-CoV-2 isolation procedure. Vero-E6 cells were infected with the positive bronchoalveolar-lavage sample. The cells were monitored daily for cytopathic effects using light microscopy.The presence of viral nucleic acid in the supernatant was detected by RT-PCR. RNA extracted from culture supernatants were used as a template to clone and sequence the genome. We used Illumina sequencing to characterize the virus genome and results showed that the isolated virus was SARS-CoV-2.

关 键 词：新型冠状病毒(SARS-CoV-2) 分离鉴定 全基因组分析 

分 类 号：R373.1[医药卫生—病原生物学]