title: New massive parallel sequencing approach improves the genetic characterization of congenital myopathies.
abstract: Congenital myopathies (CMs) are a heterogeneous group of muscle diseases characterized by hypotonia, delayed motor skills and muscle weakness with onset during the first years of life. The diagnostic workup of CM is highly dependent on the interpretation of the muscle histology, where typical pathognomonic findings are suggestive of a CM but are not necessarily gene specific. Over 20 loci have been linked to these myopathies, including three exceptionally large genes (TTN, NEB and RYR1), which are a challenge for molecular diagnosis. We developed a new approach using massive parallel sequencing (MPS) technology to simultaneously analyze 20 genes linked to CMs. Assay design was based on the Ion AmpliSeq strategy and sequencing runs were performed on an Ion PGM system. A total of 12 patients were analyzed in this study. Among the 2534 variants detected, 14 pathogenic mutations were successfully identified in the DNM2, NEB, RYR1, SEPN1 and TTN genes. Most of these had not been documented and/or fully characterized, hereby contributing to expand the CM mutational spectrum. The utility of this approach was demonstrated by the identification of mutations in 70% of the patients included in this study, which is relevant for CMs especially considering its wide phenotypic and genetic heterogeneity.
authors: Unidade de Genética Molecular, Centro de Genética Médica Dr Jacinto Magalhães, Centro Hospitalar do Porto, Porto, Portugal.|Unidade Multidisciplinar de Investigação Biomédica (UMIB), Instituto de Ciências Biomédicas Abel Salazar (ICBAS), Universidade do Porto, Porto, Portugal.
authors: Jorge
authors: J
authors: Oliveira
authors: Unidade de Genética Molecular, Centro de Genética Médica Dr Jacinto Magalhães, Centro Hospitalar do Porto, Porto, Portugal.|Unidade Multidisciplinar de Investigação Biomédica (UMIB), Instituto de Ciências Biomédicas Abel Salazar (ICBAS), Universidade do Porto, Porto, Portugal.
authors: Ana
authors: A
authors: Gonçalves
authors: Unidade de Neuropatologia, Centro Hospitalar do Porto, Porto, Portugal.
authors: Ricardo
authors: R
authors: Taipa
authors: Unidade de Neuropatologia, Centro Hospitalar do Porto, Porto, Portugal.
authors: Manuel
authors: M
authors: Melo-Pires
authors: Unidade de Genética Molecular, Centro de Genética Médica Dr Jacinto Magalhães, Centro Hospitalar do Porto, Porto, Portugal.|Unidade Multidisciplinar de Investigação Biomédica (UMIB), Instituto de Ciências Biomédicas Abel Salazar (ICBAS), Universidade do Porto, Porto, Portugal.
authors: Márcia E
authors: ME
authors: Oliveira
authors: Instituto de Patologia e Imunologia Molecular da Universidade do Porto, Porto, Portugal.|Faculdade de Medicina da Universidade do Porto, Porto, Portugal.
authors: José Luís
authors: JL
authors: Costa
authors: Instituto de Patologia e Imunologia Molecular da Universidade do Porto, Porto, Portugal.|Faculdade de Medicina da Universidade do Porto, Porto, Portugal.
authors: José Carlos
authors: JC
authors: Machado
authors: Departamento de Neurologia, Hospital Egas Moniz, Centro Hospitalar de Lisboa Ocidental, Lisboa, Portugal.
authors: Elmira
authors: E
authors: Medeiros
authors: Unidade Clínica de Paramiloidose, Centro Hospitalar do Porto, Porto, Portugal.
authors: Teresa
authors: T
authors: Coelho
authors: Consulta de Doenças Neuromusculares, Serviço de Neuropediatria, Centro Hospitalar do Porto, Porto, Portugal.
authors: Manuela
authors: M
authors: Santos
authors: Unidade de Genética Molecular, Centro de Genética Médica Dr Jacinto Magalhães, Centro Hospitalar do Porto, Porto, Portugal.|Unidade Multidisciplinar de Investigação Biomédica (UMIB), Instituto de Ciências Biomédicas Abel Salazar (ICBAS), Universidade do Porto, Porto, Portugal.|UCIBIO/REQUIMTE, Departamento de Ciências Biológicas, Laboratório de Bioquímica, Faculdade de Farmácia, Universidade do Porto, Porto, Portugal.
authors: Rosário
authors: R
authors: Santos
authors: Unidade Multidisciplinar de Investigação Biomédica (UMIB), Instituto de Ciências Biomédicas Abel Salazar (ICBAS), Universidade do Porto, Porto, Portugal.|Departamento de Microscopia, Laboratório de Biologia Celular, Instituto de Ciências Biomédicas Abel Salazar (ICBAS), Universidade do Porto, Porto, Portugal.|Centro de Genética da Reprodução Prof. Alberto Barros, Porto, Portugal.
authors: Mário
authors: M
authors: Sousa
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000483:Alleles; D019943:Amino Acid Substitution; D001706:Biopsy; D002648:Child; D004252:DNA Mutational Analysis; D034285:Dynamin II / Q000235:genetics; D005260:Female; D056726:Genetic Association Studies; D005838:Genotype; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods; D006801:Humans; D008297:Male; D008875:Middle Aged; D018482:Muscle, Skeletal / Q000473:pathology; D009135:Muscular Diseases / Q000151:congenital* / Q000175:diagnosis*; D009154:Mutation; D010375:Pedigree; D010641:Phenotype; D019837:Ryanodine Receptor Calcium Release Channel / Q000235:genetics; D055815:Young Adult
chemical_list: D019837:Ryanodine Receptor Calcium Release Channel; D034285:Dynamin II