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

Autor: Kwan Woo Kim, Sungmi Choi, Su-Kyoung Shin, Imchang Lee, Keun Bon Ku, Seong Jun Kim, Seil Kim, Hana Yi
Jazyk: angličtina
Rok vydání: 2021
Předmět:
Zdroj: Frontiers in Microbiology, Vol 12 (2021)
Druh dokumentu: article
ISSN: 1664-302X
DOI: 10.3389/fmicb.2021.602754
Popis: 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.
Databáze: Directory of Open Access Journals