摘要
为了进一步准确筛查、鉴定新型冠状病毒SARS-CoV-2,寻找可能与传播能力和症状强弱相关的变异位点,对12例意大利和西班牙国外输入浙江省的病例和7例国内病例引起的新冠肺炎临床标本进行了高通量宏转录组(metatranscriptomic)测序和基于杂交捕获的病毒基因组测序。结果表明,与宏转录组测序相比,捕获测序样本具有一致的准确突变检测结果,且所需数据量仅为宏转录组测序的1/10或更少,是一种更适合于低病毒载量样本的检测方法。SARS-CoV-2参考基因组(EPI_ISL_402119)突变分析表明,国内样本大部分都是ORF8:L84S突变株,国外样本主要是S:D614G突变株,其余样本在病毒基因ORF1a发现2种突变型,分别是ORF1a:C409S和ORF1a:P1207H。变异分析表明,在中国及意大利的新型冠状病毒肺炎患者中检测到的SARS-CoV-2病毒基因组均存在不一样的变异类群。同时,样本间检测具有差异的单核苷酸变异(SNVs),表明了SARS-CoV-2病毒变异的多态性。对同一宿主体内的全部病毒测序数据进行宿主内单核苷酸变异(iSNVs)分析表明,F06和F11体内病毒基因区的单核苷酸变异与来自同一家意大利餐厅的其他5个病例样本的SNVs突变结果意外一致,表明同一患者中可能存在重复感染。
In order to further screen and identify SARS-CoV-2 accurately and to look for mutation sites which may be re-lated to the ability of transmission and the intensity of symptoms,we have conducted metatranscriptome sequencing and hybrid capture-based sequencing for 12 imported cases confirmed COVID-19 back from Europe to Zhejiang province and 7 domestic SARS-CoV-2 infected cases from China.The result indicated that the capture sequencing samples have accurate mutation de-tection results,whereas the required data is only~1/10 or less of metatranscriptomic sequencing,which is a more suitable de-tection method for low viral load samples.Mutation analysis with reference genome(EPI_ISL_402119)showed that most of the domestic virus samples are ORF8:L84S mutation,foreign virus samples are mainly S:D614G mutation,and the rest of the samples were found 2 mutations in the virus gene ORF1a,namely ORF1a:C409S and ORF1a:P1207H.Variation analysis indi-cated that there are different mutation groups of the SARS-CoV-2 virus genomes in patients with SARS-CoV-2 between China and Italy.The detection of different single nucleotide variations(SNVs)between samples indicated the variation polymorphism of SARS-CoV-2 genomes.Analysis of intra-individual single nucleotide variations(iSNVs)of virus sequencing data in the same host indicated that the iSNVs in gene region of F06 and F11 were unexpectedly the same as the mutation of the other 5 samples from the same Italian restaurant,which indicated they might had co-infection in the same patient.
作者
曾永
陈建威
崔恩海
胡国海
王晓光
季巧英
丛迎
杨阳
杨伟斌
ZENG Yong;CHEN Jianwei;CUI Enhai;HU Guohai;WANG Xiaoguang;JI Qiaoying;CONG Ying;YANG Yang;YANG Weibin(Qingtian Center for Disease Control and Prevention,Qingtian 323900,China;BGI-Qingdao,BGI-Shenzhen,Qingdao 266555,China;Huzhou Central Hospital,Huzhou 313000,China;China National GeneBank,BGI-Shenzhen,Shenzhen 518083,China;Lishui Center for Disease Control and Prevention,Lishui 323000,China;Lishui Central Hospital,Lishui 323000,China)
出处
《激光生物学报》
2021年第4期305-315,共11页
Acta Laser Biology Sinica
基金
丽水市科技计划项目(2020KJGG10)
丽水市科技计划项目(2020KJGG07)
湖州市新冠重点研发计划项目(2020ZDT2006)
山东省重大科技创新工程项目(2020SFXGFY04-2)
国家科技重大专项项目(2017ZX10303406)。
