Agriculture Victoria Research, AgriBio Centre for AgriBioscience, 5 Ring Road, Bundoora, Victoria, 3083, Australia; School of Applied Systems Biology, La Trobe University, Bundoora, Victoria, 3086, Australia.
Agriculture Victoria Research, AgriBio Centre for AgriBioscience, 5 Ring Road, Bundoora, Victoria, 3083, Australia.
J Virol Methods. 2017 Nov;249:79-84. doi: 10.1016/j.jviromet.2017.08.019. Epub 2017 Sep 4.
With its small size and low cost, the hand-held MinION sequencer is a powerful tool for in-field surveillance. Using a metagenomic approach, it allows non-targeted detection of viruses in a sample within a few hours. This study aimed to determine the ability of the MinION to metagenomically detect and characterise a virus from an infected mosquito. RNA was extracted from an Aedes notoscriptus mosquito infected with Ross River virus (RRV), converted into cDNA and sequenced on the MinION. Bioinformatic analysis of the MinION reads led to detection of full-length RRV, with reads of up to 2.5kb contributing to the assembly. The cDNA was also sequenced on the MiSeq sequencer, and both platforms recovered the RRV genome with >98% accuracy. This proof of concept study demonstrates the metagenomic detection of an arbovirus, using the MinION, directly from a mosquito with minimal sample purification.
这款手持 MinION 测序仪体积小巧、成本低廉,是现场监测的有力工具。它采用宏基因组学方法,能够在数小时内对样本中的病毒进行非靶向检测。本研究旨在确定 MinION 能否对感染病毒的蚊子进行宏基因组检测和病毒特征分析。从感染罗斯河病毒 (RRV) 的致倦库蚊中提取 RNA,转化为 cDNA 并在 MinION 上进行测序。对 MinION 读取序列进行生物信息学分析,检测到全长的 RRV,其中长达 2.5kb 的读取序列有助于组装。cDNA 也在 MiSeq 测序仪上进行了测序,两种平台对 RRV 基因组的恢复准确率均>98%。这项概念验证研究表明,使用 MinION 可直接从经过最小程度样本纯化的蚊子中进行虫媒病毒的宏基因组检测。