Assessment of metagenomic Nanopore and Illumina sequencing for recovering whole genome sequences of chikungunya and dengue viruses directly from clinical samples
2018
Background The recent global emergence and re-emergence of arboviruses has caused significant human disease. Common vectors, symptoms and geographical distribution make differential diagnosis both important and challenging. Aim To investigate the feasibility of
metagenomicsequencing for recovering
whole genome sequencesof
chikungunyaand dengue viruses from clinical samples. Methods We performed
metagenomicsequencing using both the Illumina MiSeq and the portable Oxford Nanopore
MinIONon clinical samples which were real-time reverse transcription-PCR (qRT-PCR) positive for
chikungunya(CHIKV) or
dengue virus(DENV), two of the most important arboviruses. A total of 26 samples with a range of representative clinical Ct values were included in the study. Results Direct
metagenomicsequencing of nucleic acid extracts from serum or plasma without viral enrichment allowed for virus identification, subtype determination and elucidated complete or near-complete genomes adequate for phylogenetic analysis. One PCR-positive CHIKV sample was also found to be
coinfectedwith DENV. Conclusions This work demonstrates that
metagenomic
whole genome sequencingis feasible for the majority of CHIKV and DENV PCR-positive patient serum or plasma samples. Additionally, it explores the use of Nanopore
metagenomicsequencing for DENV and CHIKV, which can likely be applied to other RNA viruses, highlighting the applicability of this approach to
front-linepublic health and potential
portable applicationsusing the
MinION.
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