%A Wongsurawat,Thidathip %A Jenjaroenpun,Piroon %A Taylor,Mariah K. %A Lee,Jasper %A Tolardo,Aline Lavado %A Parvathareddy,Jyothi %A Kandel,Sangam %A Wadley,Taylor D. %A Kaewnapan,Bualan %A Athipanyasilp,Niracha %A Skidmore,Andrew %A Chung,Donghoon %A Chaimayo,Chutikarn %A Whitt,Michael %A Kantakamalakul,Wannee %A Sutthent,Ruengpung %A Horthongkham,Navin %A Ussery,David W. %A Jonsson,Colleen B. %A Nookaew,Intawat %D 2019 %J Frontiers in Microbiology %C %F %G English %K Native RNA,Genome,Subgenomic mRNAs,single-stranded RNA virus,Rapid detection,MinION nanopore device ®,direct RNA sequencing %Q %R 10.3389/fmicb.2019.00260 %W %L %M %P %7 %8 2019-February-25 %9 Technology Report %# %! Rapid Sequencing of RNA Viruses %* %< %T Rapid Sequencing of Multiple RNA Viruses in Their Native Form %U https://www.frontiersin.org/articles/10.3389/fmicb.2019.00260 %V 10 %0 JOURNAL ARTICLE %@ 1664-302X %X Long-read nanopore sequencing by a MinION device offers the unique possibility to directly sequence native RNA. We combined an enzymatic poly-A tailing reaction with the native RNA sequencing to (i) sequence complex population of single-stranded (ss)RNA viruses in parallel, (ii) detect genome, subgenomic mRNA/mRNA simultaneously, (iii) detect a complex transcriptomic architecture without the need for assembly, (iv) enable real-time detection. Using this protocol, positive-ssRNA, negative-ssRNA, with/without a poly(A)-tail, segmented/non-segmented genomes were mixed and sequenced in parallel. Mapping of the generated sequences on the reference genomes showed 100% length recovery with up to 97% identity. This work provides a proof of principle and the validity of this strategy, opening up a wide range of applications to study RNA viruses.