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

Kwan Woo Kim, Sungmi Choi, Su Kyoung Shin, Imchang Lee, Keun Bon Ku, Seong Jun Kim, Seil Kim, Hana Yi

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish
Article number602754
JournalFrontiers in Microbiology
Volume12
DOIs
Publication statusPublished - 2021 Feb 19

Keywords

  • coronavirus
  • MERS (Middle East respiratory syndrome)
  • MinION long-read sequencing
  • universal primer
  • whole-genome sequencing

ASJC Scopus subject areas

  • Microbiology
  • Microbiology (medical)

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