AUTHOR=Wongsurawat Thidathip , Jenjaroenpun Piroon , Taylor Mariah K. , Lee Jasper , Tolardo Aline Lavado , Parvathareddy Jyothi , Kandel Sangam , Wadley Taylor D. , Kaewnapan Bualan , Athipanyasilp Niracha , Skidmore Andrew , Chung Donghoon , Chaimayo Chutikarn , Whitt Michael , Kantakamalakul Wannee , Sutthent Ruengpung , Horthongkham Navin , Ussery David W. , Jonsson Colleen B. , Nookaew Intawat 

TITLE=Rapid Sequencing of Multiple RNA Viruses in Their Native Form

JOURNAL=Frontiers in Microbiology

VOLUME=10

YEAR=2019

URL=https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2019.00260

DOI=10.3389/fmicb.2019.00260

ISSN=1664-302X

ABSTRACT=<p>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.</p>