Comprehensive Molecular Analysis of DMD Gene Increases the Diagnostic Value of Dystrophinopathies: A Pilot Study in a Southern Italy Cohort of Patients.
Fatima Domenica Elisa De PalmaMarcella NunziatoValeria DArgenioMaria SavareseGabriella EspositoFrancesco SalvatorePublished in: Diagnostics (Basel, Switzerland) (2021)
Duchenne/Becker muscular dystrophy (DMD/BMD) is an X-linked neuromuscular disease due to pathogenic sequence variations in the dystrophin (DMD) gene, one of the largest human genes. More than 70% of DMD gene defects result from genomic rearrangements principally leading to large deletions, while the remaining are small nucleotide variants, including nonsense and missense variants, small insertions/deletions or splicing alterations. Considering the large size of the gene and the wide mutational spectrum, the comprehensive molecular diagnosis of DMD/BMD is complex and may require several laboratory methods, thus increasing the time and costs of the analysis. In an attempt to simplify DMD/BMD molecular diagnosis workflow, we tested an NGS method suitable for the detection of all the different types of genomic variations that may affect the DMD gene. Forty previously analyzed patients were enrolled in this study and re-analyzed using the next generation sequencing (NGS)-based single-step procedure. The NGS results were compared with those from multiplex ligation-dependent probe amplification (MLPA)/multiplex PCR and/or Sanger sequencing. Most of the previously identified deleted/duplicated exons and point mutations were confirmed by NGS and 1 more pathogenic point mutation (a nonsense variant) was identified. Our results show that this NGS-based strategy overcomes limitations of traditionally used methods and is easily transferable to routine diagnostic procedures, thereby increasing the diagnostic power of DMD molecular analysis.
Keyphrases
- muscular dystrophy
- duchenne muscular dystrophy
- copy number
- genome wide
- genome wide identification
- end stage renal disease
- ejection fraction
- newly diagnosed
- prognostic factors
- endothelial cells
- dna methylation
- real time pcr
- patient reported outcomes
- gene expression
- peritoneal dialysis
- transcription factor
- quantum dots
- single molecule
- minimally invasive
- living cells
- electronic health record
- nucleic acid