Abstract

Background: At the end of 2019, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) rapidly spread worldwide. Until recently, over 190 million cases and nearly four million deaths have been reported globally due to the virus. A point mutation in the SARS-CoV-2 spike protein, the D614G variant, was identified in late February 2020. Studies showed that the D614G mutation is related to higher infectivity. The current study was conducted to determine the validity of VirSNiP SARS-CoV-2 Spike D614G assay to detect the SARS-CoV-2 D614G mutation.
Methods: In this study, the detection of the SARS-CoV-2 D614G mutation was performed using VirSNiP SARS-CoV-2 Spike D614G assay (TIB Molbiol) and by Sanger sequencing. Two plasmids carrying A allele (wild-type) and G allele (mutant type) were designed using GenScript’s OptimumGene gene design tool (GenScript Inc., Piscataway, NJ, USA).
Results: It was found that VirSNiP SARS-CoV-2 Spike D614G assay could detect the SARS-CoV-2 D614G mutation. The specific mutation is easily distinguishable in the melting peaks. All of the samples were confirmed using Sanger sequencing.
Conclusions: In summary, it was shown that VirSNiP SARS-CoV-2 Spike D614G assay is a rapid, accurate, and reliable method for identifying the SARS-CoV-2 D614G mutation. In contrast, Sanger sequencing is time consuming and labor intensive. We strongly believe that VirSNiP SARS-CoV-2 Spike D614G assay will provide more information on the prevalence of the D614G mutation in Taiwanese populations.

DOI: 10.7754/Clin.Lab.2021.211009