AB043. Nanopore sequencing for genotyping Dengue virus

Abstract

Nanopore sequencer, MinION, has enabled sequencing analysis without pre-installation of expensive conventional sequencers or pre-requisite of specific skills in biological experiments. Even electric supply is not always necessary, by connecting MinION to a laptop PC. These features of MinION have opened the opportunity to enable precise genotyping of pathogens in tropical diseases on site. In this study, we attempted genotyping Dengue viruses regarding their serotypes (types 1-4). We directly used serum samples of Indonesian Dengue patients, from which viral genomes were directly amplified by the reverse-transcription-LAMP method in an isothermal reaction condition. We directly used the amplified templates for MinION sequencing allocating one flow cell per sample. We found, although the overall sequencing quality was low (80% sequence identify to the reference genome and the quality value of QV =5 on average), thereby obtained sequence data could discriminate different serotypes of the viruses, whose genome sequences were diverged with the sequence similarity of 70%, with the overall accuracy of 98%.