Laboratory of Virology, Division of Intramural Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, Montana.
Joan C. Edwards School of Medicine, Marshall University, Huntington, West Virginia.
J Infect Dis. 2018 Nov 22;218(suppl_5):S301-S304. doi: 10.1093/infdis/jiy290.
Sequencing viral genomes during an outbreak can facilitate response and containment efforts. In this study, we describe a reverse transcription long-range polymerase chain reaction for efficient amplification and sequencing of the Ebola virus (EBOV) genome in 2 seminested reactions. We demonstrate that our method remains robust with complex biological samples by amplifying and sequencing the EBOV genome from EBOV-infected nonhuman primates (NHPs). We further demonstrate that we are able to recover viral genomes from starting concentrations as low as 103 50% tissue culture infective dose (TCID50)/mL, suggesting that this method can be employed to sequence EBOV genomes from ecologically or clinically derived samples.
在疫情爆发期间对病毒基因组进行测序可以促进应对和遏制工作。在这项研究中，我们描述了一种逆转录长距离聚合酶链反应，用于在 2 次嵌套反应中有效地扩增和测序埃博拉病毒 (EBOV) 基因组。我们证明，我们的方法通过从感染 EBOV 的非人灵长类动物 (NHP) 中扩增和测序 EBOV 基因组，仍然具有复杂生物样本的稳健性。我们进一步证明，我们能够从低至 103 50%组织培养感染剂量 (TCID50)/mL 的起始浓度恢复病毒基因组，这表明该方法可用于从生态或临床衍生样本中测序 EBOV 基因组。
