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.
【저자키워드】 whole-genome sequencing, coronavirus, MERS (middle east respiratory syndrome), universal primer, MinION long-read sequencing, 【초록키워드】 public health, Mutation, protocol, Variation, Sequencing, Genome, variant, Epidemics, Diagnosis, MERS, MERS-CoV, Outbreaks, PCR, outbreak, CoV, WGS, Middle East, serum samples, sequence, syndrome, primer, whole genome, Complete, recent, CoV genome, clinical sample, Result, amplified, develop, sequenced, applied, contribute, MERS-CoV genome, 【제목키워드】 respiratory, Middle East,