Flint Annika, Reaume Spencer, Harlow Jennifer, Hoover Emily, Weedmark Kelly, Nasheri Neda
Genomics Laboratory, Bureau of Microbial Hazards, Health Canada, Ottawa, ON, Canada.
National Food Virology Reference Centre, Bureau of Microbial Hazards, Health Canada, Ottawa, ON, Canada.
Virus Evol. 2021 Sep 15;7(2):veab079. doi: 10.1093/ve/veab079. eCollection 2021.
Whole-genome sequence analysis of noroviruses is routinely performed by employing a metagenomic approach. While this methodology has several advantages, such as allowing for the examination of co-infection, it has some limitations, such as the requirement of high viral load to achieve full-length or near full-length genomic sequences. In this study, we used a pre-amplification step to obtain full-length genomic amplicons from 39 Canadian GII isolates, followed by deep sequencing on Illumina and Oxford Nanopore platforms. This approach significantly reduced the required viral titre to obtain full-genome coverage. Herein, we compared the coverage and sequences obtained by both platforms and provided an in-depth genomic analysis of the obtained sequences, including the presence of single-nucleotide variants and recombination events.
诺如病毒的全基因组序列分析通常采用宏基因组学方法进行。虽然这种方法有几个优点,比如可以检测共感染情况,但也有一些局限性,比如需要高病毒载量才能获得全长或接近全长的基因组序列。在本研究中,我们使用了一个预扩增步骤,从39株加拿大GII分离株中获得全长基因组扩增子,随后在Illumina和牛津纳米孔平台上进行深度测序。这种方法显著降低了获得全基因组覆盖所需的病毒滴度。在此,我们比较了两个平台获得的覆盖度和序列,并对获得的序列进行了深入的基因组分析,包括单核苷酸变异和重组事件的存在情况。