Metagenomic arbovirus detection using MinION nanopore sequencing

J Virol Methods. 2017 Nov:249:79-84. doi: 10.1016/j.jviromet.2017.08.019. Epub 2017 Sep 4.

Abstract

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.

Keywords: Culicidae; MiSeq; MinION; Mosquito; Ross River virus; Surveillance.

MeSH terms

  • Aedes / virology*
  • Animals
  • Computational Biology
  • DNA, Complementary
  • Genome, Viral
  • High-Throughput Nucleotide Sequencing / economics
  • High-Throughput Nucleotide Sequencing / instrumentation*
  • High-Throughput Nucleotide Sequencing / methods*
  • Metagenomics / methods*
  • Nanopores
  • Proof of Concept Study
  • Ross River virus / genetics*
  • Ross River virus / isolation & purification*
  • Sequence Analysis, DNA

Substances

  • DNA, Complementary