Table 1. Primers used in this study.
| Gene/Exon | Forward Primer | Reverse Primer | Amplicon size (bp) | Multiplex set | |
| qPCR-HRM1 | |||||
| B2M/2 | cactgaaaaagatgagtatgcc | Aacattccctgacaatccc | 231 | ||
| DMD/16 | tctatgcaaatgagcaaatacacgc | ggtatcactaacctgtgctgtactc | 290 | ||
| MVK/2 | tcttcctgctggttctgaca | gcatcagggagaaggggta | 203 | ||
| MVK/3 | tcaccctcaggcttattgct | tggtttctcctccttgcact | 210 | ||
| MVK/4 | ccctctcacccacttgtgtt | aggctgaatctggactccttc | 226 | ||
| MVK/5 | cgggagagtcacgtttcac | gacactggccaggtaaggac | 221 | ||
| MVK/6 | ccactcctcactgccacag | ctcttgggcacctaccattg | 221 | ||
| MVK/7 | cctctctcccaagtagcacag | ccctgcacacatcaatgc | 170 | ||
| MVK/8 | tgagttcagtgtggacctgc | taattgtgtcctggccttcc | 181 | ||
| MVK/92 | gaacacctcctccctccac | ttctgagcacagccagattg | 295 | ||
| MVK/10 | aagtgggaacagatggaacct | ccaatgaggaagcaaagacc | 346 | ||
| MVK/11 | gtcaagggtgacctgccttc | gcctctccagcagtgtcag | 300 | ||
| NLRP3/3a | gatgtgtgtatactttccccctaa | aaacctgtcttggtagagtgtcc | 399 | ||
| NLRP3/3c | agcctcatcagaaagaagctg | tgaggtcggactcctcaaac | 469 | ||
| NLRP3/3d | tctttggctgcagatggaat | ttgctgagagatcttgcaactta | 399 | ||
| NLRP3/3e | tttcctctttggcctggtaa | agaagaagctggcgaggaag | 477 | ||
| NLRP3/3c3 | gcctctctgctcatcacgac | gctcttgccactctccatct | 264 | ||
| TNFRSF1A/2 | ttgatggtgtctcctctatctga | aagaagcagcaccccagac | 240 | ||
| TNFRSF1A/3 | gggctccttccttgtgttct | cacatagacaggcacccaca | 247 | ||
| TNFRSF1A/4 | agaaatgggtcaggtggagat | gccagagaggagttggttgt | 280 | ||
| qPCR | |||||
| MVK/92 | accagccgttccttcttttt | ttctgagcacagccagattg | 230 | ||
| NLRP3/2 | ggggtctcctctctcatgc | ctagaagcaccaccccagtc | 242 | ||
| NLRP3/4 | tcgaggctgatttcttttctg | gcactcacacagatcacatgc | 248 | ||
| NLRP3/5 | tctgatgctttctgcctctgt | cgctggcagaacttccttag | 244 | ||
| NLRP3/6 | tgactgacattctgccatctct | tctggtaagacacccatgaaga | 227 | ||
| NLRP3/7 | ggaacagctgggtactgagg | cttgtggccactctgccta | 295 | ||
| NLRP3/8 | aatcaccccctttttgcag | agagccatcctggattttga | 274 | ||
| NLRP3/9a | agtgcaacccaggctttcta | tcacagagctgtggtcttgg | 240 | ||
| SQF-PCR4 | |||||
| GFAP/3 | gaggaaaggattgatggcca | gaggaggagatccggttctt | 249 | Control (A to H) | |
| DMD/28 | tgcattttgaattacctgctaca | agtaccaaatagaagacaaatccaaag | 361 | Control (A to H) | |
| MVK/2 | tattatgatgggcttgaactagg | tgcctcagggtgtcctttta | 292 | A | |
| MVK/3 | cttcttagcacgtgggtcct | ctctctgtaggctctt | 286 | A | |
| MVK/4 | gtcgattttctgtgttctgttgtt | agagcatgtgcattctccag | 337 | B | |
| MVK/5 | ctggaccagatgcttggagt | aagccacgtccctgtcctg | 336 | A | |
| MVK/6 | gagtggacttgttctttctgagc | agactcttgggcacctacca | 328 | B | |
| MVK/7 | ttcctgaatggggcaaaat | ctgcctcctatggtacttccc | 266 | C | |
| MVK/8 | gtatcagggtgggcggcttcc | gagggagacctcgaaaatcc | 279 | A | |
| MVK/9 | ggctgtgtgaacacctcctc | cacagccagattgcagagcca | 314 | B | |
| MVK/10 | tctccagccaacaactgtca | tcaagggaattctccaggtg | 348 | C | |
| MVK/11 | ctgggcttttgccttgaat | aataatccagaaaggggcatc | 304 | C | |
| TNFRSF1A/1 | accaggccgtgatctctatg | cactcttccctttgtccctg | 224 | H | |
| TNFRSF1A/2 | aggacttgagccagggaagt | tttccttggggacacacact | 198 | D | |
| TNFRSF1A/3 | ctggctgttgtccctagcat | cacccacacaccactcaaga | 256 | E | |
| TNFRSF1A/4 | agaaatgggtcaggtggagat | ttggttgtcagacccacaga | 268 | D | |
| TNFRSF1A/5 | acaaccaacttcctctctggc | atctgttgcccagctaatgg | 277 | I | |
| TNFRSF1A/6 | caccagtgccgtctcttctt | atagatggatgggtgggatg | 186 | I | |
| TNFRSF1A/7 | aacacctgctttgtctgcag | accttctgcccagagtccc | 268 | H | |
| TNFRSF1A/8–9 | gggaaatcgacacctgaaaa | aagctccccctgaaagagag | 233 | H | |
| TNFRSF1A/9–10 | cccttcagaagtgggaggac | gatcgatctcgtggtcgct | 305 | I | |
| NLRP3/1 | ccagagccttcagtttggag | aggagtgtgtcctgagccat | 269 | G | |
| NLRP3/2 | ccactgtgatatgccaggaa | gcattccaaagagcaggaac | 208 | F | |
| NLRP3/3 | gcatctcaggtggatgtgtg | caggctcagaatgctcatca | 307 | G | |
| NLRP3/4 | cctcacttccagtttttgcc | gcactcacacagatcacatgc | 189 | G | |
| NLRP3/5 | tctgtgtgtgggactgaagc | ctttccccacgacaaacact | 228 | F | |
| NLRP3/6 | tcagtattgagcaccagcca | tgaccaaagtaacccccatc | 213 | G | |
| NLRP3/7 | gagtcaaagcagctgcacaa | ccaccatgtgttctcattgc | 283 | F | |
| NLRP3/8 | gggatggttaaggggacatt | caggcccaacctaatcttga | 339 | G | |
| NLRP3/9a | tgcaacccaggctttctatt | tgctgtcattgtcctggtgt | 348 | F |
1
All primers (except HRM-qPCR MVK/9 primer) showed a PCR efficiency range between 80 and 100% (IC+/−0.2%).
2
qPCR and HRM of MVK exon 9 could not be performed in the same run due to low PCR efficiency. We used therefore two different primer pairs.
3
p.T348M allele specific primer.
4
Forward primers were 6FAM labeled. Nine distinct PCR sets (Multiplex A to I) were designed, each yielding a pattern of 3 to 6 peaks including the 2 control fragments (DMD and GFAP).