Table 4. Validation of NGS for variant/mutation detection on known CMD-positives samples.
| Sample | Gene | Mutations/variants detected by Sanger sequencing | Mutation/variant type | Detected by NGS | Coverage | Mutant allele % |
| CMD-8 | COL6A1 | c.1931G>A (p.R644Q), Het | missense | − | 11 | - |
| COL6A2 | c.1770G>C (p.T590), Het | silent | + | 8 | 25 | |
| COL6A2 | c.2994C>T (p.H998), Het | silent | + | 20 | 50 | |
| CMD-9 | COL6A1 | IVS29-8G>A, Het | intronic | + | 7 | 71 |
| FKTN | IVS9-40C>A, Het | intronic | (−) | - | - | |
| LAMA2 | c.2084C>T (p.D695V), Het | missense | + | 82 | 52 | |
| LAMA2 | c.5614G>T (p.D1872Y), Het | missense | + | 29 | 31 | |
| SEPN1 | IVS5+39C>T, Het | intronic | (−) | - | - | |
| SEPN1 | IVS11-31C>T, Het | intronic | (−) | - | - | |
| SEPN1 | c.1645G>A (p.V549M), Het | missense | + | 78 | 38 | |
| SEPN1 | c.1773+44G>T, Het | 3′ UTR | − | - | - | |
| CMD-10 | COL6A1 | IVS26+50C>T, Het | intronic | (−) | - | - |
| COL6A1 | c.2424G>T (p.Q808H), Het | missense | − | 17 | - | |
| COL6A2 | IVS24-3dupC, Het | duplication | − | 8 | - | |
| COL6A3 | IVS38-34C>T, Homo | intronic | (−) | - | - | |
| CMD-11 | LAMA2 | c.3154A>G, Het | splicing | + | 17 | 58 |
| LAMA2 | c.6617delT, Het | deletion | − | 1 | - | |
| CMD-12 | POMGNT1 | c.636C>T (p.F212), Het | splicinga | + | 34 | 29 |
| POMGNT1 | IVS17+1G>A, Het | splicinga | + | 105 | 55 | |
| CMD-13 | - | - | - | − | - | - |
a
This mutation has been reported in individuals with MEB disease; +, detected; −, Not detected; (−) Mutation and/or variant not detected because the bioinformatics algorithm for NGS data is set to detect +/−20 bases from exon/intron boundaries.