TABLE 1.
Details of the four assays used in the multiplex HRM method
| Assay | Target | Primer | Sequence (5′–3′) | Vol (μL)a |
|---|---|---|---|---|
| Assay 1 | N | N-F | AGTCACACCTTCGGGAACG | 1 |
| N-R | AGGCCTGAGTTGAGTCAGCA | 1 | ||
| S | G142D-F | CTGTGAATTTCAATTTTGTAATGATCCATT | 0.5 | |
| G142D-R | CAACTTTTGTTGTTTTTGTGGTAATAAACA | 0.5 | ||
| S | R158G-F | AAAGTTGGATGGAAAGTGAGTTC | 1 | |
| R158G-R | AGGCTGAGAGACATATTCAAAAGTG | 1 | ||
| Assay 2 | N | 1 | ||
| 1 | ||||
| S | Y505H-F | TCCAACCCACTAATGGTGTTG | 1 | |
| Y505H-R | AGAAAGTACTACTACTCTGTATGGT | 1 | ||
| S | V213G-F | CTAAGCACACGCCTATTAATTTAG | 1 | |
| V213G-R | CCTGAGGGAGATCACGC | 1 | ||
| Assay 3 | N | 1 | ||
| 1 | ||||
| S | G446S-F | ATTCTAACAATCTTGATTCTAAGGTT | 0.5 | |
| G446S-R | CAATCTATACAGGTAATTATAATTACCAC | 0.5 | ||
| N | S413R-F | CTGCTGCAGATTTGGATGATT | 0.5 | |
| S413R-R | TTAGGCCTGAGTTGAGTCAG | 0.5 | ||
| Assay 4 | N | 1 | ||
| 1 | ||||
| S | F486V-F | ATCTTGATTCTAAGGTTGGTGGTAATTATA | 0.5 | |
| F486V-R | GGTTGGAAACCATATGATTGTAAAGGAAA | 0.5 | ||
| S | S704L-F | GGCGGGCACGTAGTGTAG | 1 | |
| S704L-R | TAGACACTGGTAGAATTTCTGTGGTA | 1 | ||
a
Volume of each 10 μM primer added to the primer pool.