A high-resolution melt curve toolkit to identify lineage-defining SARS-CoV-2 mutations

Sci Rep. 2023 Mar 8;13(1):3887. doi: 10.1038/s41598-023-30754-1.

Abstract

The emergence of severe acute respiratory syndrome 2 (SARS-CoV-2) variants of concern (VOCs), with mutations linked to increased transmissibility, vaccine escape and virulence, has necessitated the widespread genomic surveillance of SARS-CoV-2. This has placed a strain on global sequencing capacity, especially in areas lacking the resources for large scale sequencing activities. Here we have developed three separate multiplex high-resolution melting assays to enable the identification of Alpha, Beta, Delta and Omicron VOCs. The assays were evaluated against whole genome sequencing on upper-respiratory swab samples collected during the Alpha, Delta and Omicron [BA.1] waves of the UK pandemic. The sensitivities of the eight individual primer sets were all 100%, and specificity ranged from 94.6 to 100%. The multiplex HRM assays have potential as a tool for high throughput surveillance of SARS-CoV-2 VOCs, particularly in areas with limited genomics facilities.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biological Assay
  • COVID-19*
  • Genomics
  • Humans
  • Mutation
  • SARS-CoV-2 / genetics