Poster Presentation 9th Australasian Virology Society Meeting 2017

Metagenomic arbovirus detection from mosquitoes using MinION sequencing (#157)

Jana Batovska 1 2 , Stacey E Lynch 1 , Brendan C Rodoni 1 2 , Tim I Sawbridge 1 2 , Noel OI Cogan 1 2
  1. Agriculture Victoria Research, AgriBio Centre for AgriBioscience, Bundoora, Victoria, Australia
  2. School of Applied Systems Biology, La Trobe University, Bundoora, Victoria, Australia

The hand-held MinION sequencer allows target-independent identification of pathogens in-field within a short time frame. The portability and affordability of this technology make it suitable for biosecurity surveillance. This study aimed to determine if the MinION could be used to detect an arbovirus from a mosquito using a metagenomic approach. The cDNA from an Aedes notoscriptus mosquito infected with Ross River virus (RRV) was sequenced on the MinION, and also on the MiSeq for comparison. Analysis of the MinION data led to the assembly of the full-length RRV genome, with 12% of the RRV reads produced within the first 10 minutes of the run, and reads of up to 2.5 kb contributing to the assembly. The MinION had comparable performance to the MiSeq, with both platforms producing RRV consensus sequences with >98% accuracy. Due to the metagenomic nature of the sequencing, the MinION data could also be used to identify the mosquito species, further demonstrating its utility in biosecurity surveillance. This study shows the MinION has sufficient sensitivity and accuracy to detect and characterise an arbovirus directly from a mosquito. With recent advancements in technology allowing for shorter sample preparation times and the mobilisation of laboratory techniques, the MinION has the potential to transform in-field surveillance.