Method: SARS-CoV-2 TaqMan Assays for
S substitutions
K417T,
E484K,
E484Q, and
L452R were performed per manufacturer's instructions (Thermo Fisher) with 4 muL of template.
Result: Amplification failure was expected and occurred for strains lacking both reference and mutation sequences at the locus (e.g., B.1.351 lacks both alleles at
K417T, B.1.351 and P.1 lack both alleles at
E484Q, and B.1.617.1 lacks both alleles at
E484K) which indicates high specificity of all assays performed on synthetic RNA.
Result: Furthermore, of the 96 replicates that produced an inconclusive result, 74 were due to the presence of an alternate allele: 70 replicates containing
E484K (B.1.351 an