新型冠状病毒南非B.1.351变异株的分离与分子特征分析  

Isolation and Characterization of“South African”SARS-CoV-2 Strain B.1.351

作　　者：郑焕英[1] 张欢[1] 陆靖 欧阳方竹 邹丽容[1] 胡瑶 李振翠 黎薇[1] 李柏生[1] 何剑锋[1] 柯昌文[1] 邓小玲[1] ZHENG Huanying;ZHANG Huan;LU Jing;OUYANG Fangzhu;ZHOU Lirong;HU Yao;LI Zhencui;LI Wei;LI Baisheng;HE Jianfeng;KE Changwen;DENG Xiaoling(Guangdong Provincial Center for Disease Control and Prevention,Guangzhou 510515,China;Guangdong Provincial Institute of Public Health,Guangzhou 511430,China)

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

出　　处：《病毒学报》2021年第3期534-540,共7页Chinese Journal of Virology

基　　金：国家重点研发计划(项目编号:2021YFC0863300),题目:新型冠状病毒变异监测及功能影响分析。

摘　　要：2020年12月广州市第八人民医院收治了1例南非输入性COVID-19病例,经检测为SARS-CoV-2核酸阳性的实验室确诊病例。本研究使用Vero-E6细胞,对该病例咽拭子样本进行病毒分离,逐日观察,咽拭子接种的细胞管3d开始出现细胞融合样细胞病变(Cytopathic effect,CPE),5d出现完全CPE后再进行第二代接种,2d出现病变,提取核酸进行鉴定,为SARS-CoV-2阳性。第2代复传的SARS-CoV-2病毒株TCID50测定结果为5.5log TCID50/0.1mL~5.8log TCID50/0.1mL。对分离毒株经采用三代测序成功获得全基因组序列,进化分析结果为与参考毒株Wuhan/Hu-1/2019相比共发生30个碱基的变异、18个氨基酸的突变和18个碱基的缺失,比对结果显示分离到的毒株为SARS-CoV-2南非B.1.351变异株。In December 2020,an imported COVID-19 case from South Africa was laboratory-confirmed and the patient was transferred to Guangzhou Eighth People’s Hospital.A clinical sample was sent to Guangdong provincial CDC for viral isolation and whole-genome sequencing.Vero-E6 cells were used to isolate the virus from oropharyngeal swab samples.Cytopathic effects(CPE)began to appear in cell tubes inoculated with the oropharyngeal swab on the third day.After complete CPE appeared on the fifth day,a second generation was inoculated,and CPE appeared on the second day.Nucleic acids were extracted for RT-PCR confirmation and sequencing.The TCID50 of the SARS-CoV-2 virus strain was 5.5 log/0.1 mL–5.8 log/0.1 mL.The wholegenome sequence was obtained by Nanopore sequencing.Evolutionary analysis showed that the isolated strain belongs to the SARS-CoV-2 B.1.351("South African")lineage.Compared with the reference strain Wuhan/Hu-1/2019,there are 30 nucleotide(nt)substitutions(18 amino acid substitutions)and 18 nt deletions in this isolate.

关 键 词：新型冠状病毒(SARS-CoV-2) 病毒分离 VERO-E6细胞 全基因组序列 

分 类 号：R373.9[医药卫生—病原生物学]