Genetic Profiling of Neuromuscular Disorders

This research was conducted in accordance with the Declaration of Helsinki. The descriptive, cross-sectional study was performed on paediatric patients (age ≤ 18 years) with MD and CM from Siriraj Neuromuscular Disease Center (Siriraj Hospital) and King Chulalongkorn Memorial Hospital in Thailand between September 2018 and April 2021. The diagnoses of MD and CM were made by paediatric neurologists and based on the presence of muscle weakness, age at onset, clinical course, physical findings, conventional investigations and muscle histopathological findings. Muscle biopsies were performed using standard immunocytochemistry protocols. Staining for dystrophin, utrophin, merosin, dysferlin, caveolin-3, dystroglycan, sarcoglycan, emerin, collagen VI and desmin was performed. Single-gene molecular tests were performed on patients with suggestive clinical and pathological findings. The molecular tests were multiplex ligation-dependent probe amplification (MLPA) for spinal muscular atrophy (SMN1) and DMD (DMD); Sanger sequencing of coding regions of RYR1 (8 out of 106 exons) and ACTA1; and mitochondrial DNA sequencing (A3243G, A8344G, T8993G). Patients suspected of having facioscapulohumeral MD (FSHD) underwent long-read DNA sequencing. Patients whose mutations were still elusive were subjected to ES (Fig. 1).Figure 1

Patient enrolment workflow.