KU Leuven, Rega Institute, Department of Microbiology, Immunology and Transplantation, Laboratory of Clinical and Epidemiological Virology, BE-3000 Leuven, Belgium.
KU Leuven, Rega Institute, Department of Microbiology, Immunology and Transplantation, Laboratory of Clinical and Epidemiological Virology, BE-3000 Leuven, Belgium.
Infect Genet Evol. 2020 Jul;81:104267. doi: 10.1016/j.meegid.2020.104267. Epub 2020 Feb 27.
Because of the enormous variation in their genome sequence, genotyping enteroviruses by standard methods can prove to be quite challenging. Nanopore sequencing offers the potential to overcome the limitations of older techniques, but thus far, only amplicon-based strategies have been used to sequence complete enterovirus genomes. By combining a sequence-independent, single primer amplification (SISPA) for cDNA generation with next-generation sequencing using the Oxford Nanopore MinION, complete enterovirus genomes can be obtained in an easy-to-use, sequence-independent manner. To demonstrate its usability, we applied this technique to determine the complete genome sequence of an enterovirus C104 strain, representing the first documented occurrence of this uncommon enterovirus strain in Belgium.
由于其基因组序列的巨大差异,通过标准方法对肠道病毒进行基因分型可能极具挑战性。纳米孔测序提供了克服旧技术局限性的潜力,但迄今为止,仅基于扩增子的策略已被用于测序完整的肠道病毒基因组。通过将 cDNA 生成的无序列依赖性单引物扩增 (SISPA) 与使用 Oxford Nanopore MinION 的下一代测序相结合,可以以一种易于使用、无序列依赖性的方式获得完整的肠道病毒基因组。为了证明其可用性,我们应用该技术来确定肠道病毒 C104 株的完整基因组序列,这是该罕见肠道病毒株在比利时首次有记录的发生。