Ann Clin Microbiol.  2021 Dec;24(4):135-140. 10.5145/ACM.2021.24.4.4.

A Case of Whole Genome Analysis of SARSCoV-2 Using Oxford Nanopore MinION System

• Kim JS
• Chung SH
• Kim JM
• Kim HS
• Kim HS
• Song W

• KMID: 2523568
• DOI: http://doi.org/10.5145/ACM.2021.24.4.4

Abstract

The application of whole genome sequencing on SARS-CoV-2 viral genome is essential for our understanding of the molecular epidemiology and spread of viruses in the community. The portable whole genome sequencer MinION (Oxford Nanopore Technologies, ONT, UK) could be feasibly used in a clinical microbiology laboratory without the need of vast resources or stringent operating conditions. We used the MinION sequencer to analyze the viral genome sequence of one SARS-CoV-2 strain. In June 2020, nasopharyngeal specimen from one patient was subjected to whole-genome analysis using the nCoV-2019 sequencing protocol v2 of ARTIC using the MinION sequencer. The ONT MinKNOW software, RAMPART tool, and Genome Workbench were used. We identified 11 nucleotide variants using the Wuhan-Hu-1 isolate (NC_045512.2) as the reference sequence. There were six nucleotide variants (T265I, F924, Y3884L, P4715L, L5462, and Q6804L) in the ORF1ab region, one variant (D614G) in the S gene, one variant (Q57H) in ORF3a, one variant (P302) in the N gene, and two variants in each the 5′-UTR and 3′-UTR. In this prolonged coronavirus disease 2019 (COVID-19) pandemic season, the MinION system that operates an amplicon-based whole-genome sequencing protocol could be a rapid and reliable sequencer without the need of cumbersome viral cultivation.

Keyword

Nanopore sequencing; SARS-CoV-2; COVID-19; Whole genome sequencing

Figure

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  Fig. 1. Graphic sequence view of NCBI Genome Workbench for D614G variant in S gene. MinION sequencer is known as a next-generation sequencer with less accuracy, but it revealed sufficient accuracy for the mutation with amplicon-based approach. NCBI, The National Center for Biotechnology Information

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