Rapid, Sensitive, Full-Genome Sequencing of Severe Acute Respiratory Syndrome Coronavirus 2

Emerg Infect Dis. 2020 Oct;26(10):2401-2405. doi: 10.3201/eid2610.201800. Epub 2020 Jul 1.

Abstract

We describe validated protocols for generating high-quality, full-length severe acute respiratory syndrome coronavirus 2 genomes from primary samples. One protocol uses multiplex reverse transcription PCR, followed by MinION or MiSeq sequencing; the other uses singleplex, nested reverse transcription PCR and Sanger sequencing. These protocols enable sensitive virus sequencing in different laboratory environments.

Keywords: COVID-19; SARS-CoV-2; coronavirus; coronavirus disease; genomics; high-throughput nucleotide sequencing; respiratory infections; severe acute respiratory syndrome coronavirus 2; viruses; whole-genome sequencing; zoonoses.

MeSH terms

  • Betacoronavirus / genetics*
  • COVID-19
  • Coronavirus Infections / virology*
  • Multiplex Polymerase Chain Reaction
  • Pandemics
  • Pneumonia, Viral / virology*
  • RNA, Viral / analysis*
  • SARS-CoV-2
  • Sequence Analysis, RNA / methods*
  • Whole Genome Sequencing / methods*

Substances

  • RNA, Viral