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通过比较MinION和Illumina iSeq100系统基于直接扩增子测序的SARS-CoV-2快速基因组特征分析

Rapid Genomic Characterization of SARS-CoV-2 by Direct Amplicon-Based Sequencing Through Comparison of MinION and Illumina iSeq100 System.

作者信息

Hourdel Véronique, Kwasiborski Aurelia, Balière Charlotte, Matheus Séverine, Batéjat Christophe Frédéric, Manuguerra Jean-Claude, Vanhomwegen Jessica, Caro Valérie

机构信息

Environment and Infectious Risks Unit, Institut Pasteur, Paris, France.

出版信息

Front Microbiol. 2020 Sep 25;11:571328. doi: 10.3389/fmicb.2020.571328. eCollection 2020.

Abstract

Global human health is increasingly challenged by emerging viral threats, especially those observed over the last 20 years with coronavirus-related human diseases, such as the Severe Acute Respiratory Syndrome (SARS) and the Middle East Respiratory Syndrome (MERS). Recently, in late December 2019, a novel , SARS-CoV-2, originating from the Chinese city of Wuhan, emerged and was then identified as the causative agent of a new severe form of pneumonia, COVID-19. Real-time genome sequencing in such viral outbreaks is a key issue to confirm identification and characterization of the involved pathogen and to help establish public health measures. Here, we implemented an amplicon-based sequencing approach combined with easily deployable next-generation sequencers, the small and hand-held MinION sequencer and the latest most compact Illumina sequencer, the iSeq100 system. Our results highlighted the great potential of the amplicon-based approach to obtain consensus genomes of SARS-CoV-2 from clinical samples in just a few hours. Both these mobile next-generation sequencers are proven to be efficient to obtain viral sequences and easy to implement, with a minimal laboratory environment requirement, providing useful opportunities in the field and in remote areas.

摘要

全球人类健康日益受到新出现的病毒威胁的挑战,尤其是在过去20年中观察到的与冠状病毒相关的人类疾病,如严重急性呼吸综合征(SARS)和中东呼吸综合征(MERS)。最近,在2019年12月底,一种源自中国武汉市的新型冠状病毒SARS-CoV-2出现了,随后被确定为一种新型严重肺炎COVID-19的病原体。在这种病毒爆发中进行实时基因组测序是确认所涉病原体的鉴定和特征以及帮助制定公共卫生措施的关键问题。在此,我们实施了一种基于扩增子的测序方法,并结合了易于部署的下一代测序仪,即小型手持式MinION测序仪和最新、最紧凑的Illumina测序仪iSeq100系统。我们的结果突出了基于扩增子的方法在短短几小时内从临床样本中获得SARS-CoV-2共识基因组的巨大潜力。事实证明,这两种便携式下一代测序仪都能有效地获取病毒序列,且易于实施,对实验室环境要求极低,为现场和偏远地区提供了有用的机会。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/eac8/7546329/6f867f4c1099/fmicb-11-571328-g001.jpg

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