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A Half-Day Genome Sequencing Protocol for Middle East Respiratory Syndrome Coronavirus

Authors
Kim, Kwan WooChoi, SungmiShin, Su-KyoungLee, ImchangKu, Keun BonKim, Seong JunKim, SeilYi, Hana
Issue Date
19-2월-2021
Publisher
FRONTIERS MEDIA SA
Keywords
MERS (Middle East respiratory syndrome); coronavirus; universal primer; whole-genome sequencing; MinION long-read sequencing
Citation
FRONTIERS IN MICROBIOLOGY, v.12
Indexed
SCIE
SCOPUS
Journal Title
FRONTIERS IN MICROBIOLOGY
Volume
12
URI
https://scholar.korea.ac.kr/handle/2021.sw.korea/129183
DOI
10.3389/fmicb.2021.602754
ISSN
1664-302X
Abstract
Recent coronavirus (CoV) outbreaks, including that of Middle East respiratory syndrome (MERS), have presented a threat to public health worldwide. A primary concern in these outbreaks is the extent of mutations in the CoV, and the content of viral variation that can be determined only by whole genome sequencing (WGS). We aimed to develop a time efficient WGS protocol, using universal primers spanning the entire MERS-CoV genome. MERS and synthetic Neoromicia capensis bat CoV genomes were successfully amplified using our developed PCR primer set and sequenced with MinION. All experimental and analytical processes took 6 h to complete and were also applied to synthetic animal serum samples, wherein the MERS-CoV genome sequence was completely recovered. Results showed that the complete genome of MERS-CoV and related variants could be directly obtained from clinical samples within half a day. Consequently, this method will contribute to rapid MERS diagnosis, particularly in future CoV epidemics.
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보건과학대학 (바이오시스템의과학부)
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