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Sequencing the Complete Genome of COVID-19 Virus from Clinical Samples Using the Sanger Method

Roujian Lu1, Peihua Niu1, Li Zhao1

  • 1Key Laboratory of Biosafety, National Health and Family Planning Commission, National Institute for Viral Disease Control and Prevention, China CDC, Beijing, China.

China CDC Weekly
|October 1, 2021
PubMed

Insights

Sanger sequencing provides a cost-effective method for obtaining complete severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) genomes from clinical samples. This technique enables the monitoring of viral genetic variations in COVID-19 infections without expensive equipment.

Area of Science:

  • Virology
  • Genomics
  • Molecular Biology

Background:

  • Coronavirus disease 2019 (COVID-19) is caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2).
  • Complete SARS-CoV-2 genomes have been previously sequenced using next-generation sequencing (NGS).

Purpose of the Study:

  • To provide technical data for sequencing complete SARS-CoV-2 genomes from clinical samples using the Sanger sequencing method.
  • To demonstrate the feasibility of using Sanger sequencing for viral genome analysis.

Main Methods:

  • Sanger sequencing was employed to obtain complete genomes of SARS-CoV-2 from clinical samples.
  • Two complete SARS-CoV-2 genomes (WH19004-S and GX0002) were successfully sequenced.
  • Single nucleotide polymorphisms (SNPs) were identified in specific regions of the WH19004-S genome.

Main Results:

  • Complete genome sequences of SARS-CoV-2 were obtained using the Sanger method.
  • Two specific single nucleotide polymorphisms (SNPs) were identified in the ORF7a and ORF8 regions of the WH19004-S genome.
  • The obtained sequences were of high quality.

Conclusions:

  • Sanger sequencing is a viable and cost-effective alternative to NGS for complete SARS-CoV-2 genome sequencing.
  • This method can generate high-quality data for monitoring viral genetic variations in COVID-19.
  • It supports public health surveillance of SARS-CoV-2 without the need for expensive equipment.

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