Rapid sequencing of multiple RNA viruses in their native form

Thidathip WongsurawatPiroon JenjaroenpunMariah K. TaylorJasper LeeAline Lavado TolardoJyothi ParvathareddySangam KandelTaylor D. WadleyBualan KaewnapanNiracha AthipanyasilpAndrew SkidmoreDonghoon ChungChutikarn ChaimayoMichael WhittWannee KantakamalakulRuengpung SutthentNavin HorthongkhamDavid W. UsseryColleen B. JonssonIntawat NookaewUniversity of Arkansas for Medical SciencesUniversity of LouisvilleFaculty of Medicine, Siriraj Hospital, Mahidol UniversityUniversidade de Sao Paulo - USPUniversity of Tennessee Health Science CenterUniversity of Arkansas at Little Rock2020-01-272020-01-272019-01-01Frontiers in Microbiology. Vol.10, No.FEB (2019)1664302X2-s2.0-85064222600https://repository.li.mahidol.ac.th/handle/20.500.14594/51161© 2019 Frontiers Media S.A. All Rights Reserved. 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.Mahidol UniversityImmunology and MicrobiologyRapid sequencing of multiple RNA viruses in their native formArticleSCOPUS10.3389/fmicb.2019.00260