福建省新型冠状病毒的分离和全基因组特征分析

Isolation and genome sequence analyses of 2019-nCoV from COVID-19 patients in Fujian Province,China

目的从福建省早期新型冠状病毒肺炎(COVID-19)病例中分离新型冠状病毒(2019-nCoV)并了解病毒基因组特征。方法将福建省早期COVID-19患者的咽拭子样本接种至Vero-E6细胞,培养上清提取核酸,经real-time RT-PCR法验证后,利用Ion Torrent S5深度测序仪做全基因组测序,分析病毒株全基因组序列与参考序列的一致性和进化情况。结果12份咽拭子样本经培养后有2份样本病毒滴度明显上升,表明分离到2株2019-nCoV,其全基因组结构蛋白与Wuhan-Hu-1株的一致性超过99.9%。结论成功自福建省早期COVID-19病例中分离到2019-nCoV,病毒分离株的基因序列与湖北武汉流行的2019-nCoV高度同源,提示病毒尚未发生明显变异。

To explore genomic characteriation of 2019-nCoV,throat swab specimens from COVID-19 patients were inoculated on Vero-E6 cells.Six days post-inoculation,supernatants of cell cultures were collected,tested,and sequenced on the Ion Torrent S5 Next-Generation Sequencing system.Two strains viruses were isolated from 12 specimens(eight of them were positive in ORF1 ab gene and N gene,one of them was positive in N gene and three of them were negative in ORF1 ab gene and N gene),as confirmed by online BLAST search at NCBI website.Genetic similarity of more than 99.9%were observed,from genomic level to structure genes including E,M,N and S genes,between the 2 isolates and the 2019-nCoV reference strain Wuhan-Hu-1 that isolated from Wuhan,Hubei province.Therefore,these results indicated that 2019-nCoV isolates in Fujian province have not mutated significantly yet so far.