Assessment of a multiplex PCR and Nanopore-based method for dengue virus sequencing in Indonesia.
Samuel C B StubbsBarbara A BlacklawsBenediktus YohanFrilasita A YudhaputriRahma F HayatiBrian SchwemEdsel M SalvañaRaul V DesturaJames S LesterKhin S MyintR Tedjo SasmonoSimon D W FrostPublished in: Virology journal (2020)
The multiplex, short-amplicon approach proved superior for amplifying DENV genomes from clinical samples, particularly when the virus was present at low concentrations. The accuracy of Nanopore-generated consensus sequences from these amplicons was sufficient for identifying the geographic origin of the samples, demonstrating that the approach can be a useful tool for identifying and monitoring DENV clades circulating in low-resource settings across Indonesia. However, the inaccuracies in Nanopore-generated consensus sequences mean that the approach may not be appropriate for higher resolution transmission studies, particularly when more accurate sequencing technologies are available.