title: Targeted Re-Sequencing Emulsion PCR Panel for Myopathies: Results in 94 Cases.
abstract: Molecular diagnostics in the genetic myopathies often requires testing of the largest and most complex transcript units in the human genome (DMD, TTN, NEB). Iteratively targeting single genes for sequencing has traditionally entailed high costs and long turnaround times. Exome sequencing has begun to supplant single targeted genes, but there are concerns regarding coverage and needed depth of the very large and complex genes that frequently cause myopathies.
  
  
  
  To evaluate efficiency of next-generation sequencing technologies to provide molecular diagnostics for patients with previously undiagnosed myopathies.
  
  
  
  We tested a targeted re-sequencing approach, using a 45 gene emulsion PCR myopathy panel, with subsequent sequencing on the Illumina platform in 94 undiagnosed patients. We compared the targeted re-sequencing approach to exome sequencing for 10 of these patients studied.
  
  
  
  We detected likely pathogenic mutations in 33 out of 94 patients with a molecular diagnostic rate of approximately 35%. The remaining patients showed variants of unknown significance (35/94 patients) or no mutations detected in the 45 genes tested (26/94 patients). Mutation detection rates for targeted re-sequencing vs. whole exome were similar in both methods; however exome sequencing showed better distribution of reads and fewer exon dropouts.
  
  
  
  Given that costs of highly parallel re-sequencing and whole exome sequencing are similar, and that exome sequencing now takes considerably less laboratory processing time than targeted re-sequencing, we recommend exome sequencing as the standard approach for molecular diagnostics of myopathies.
authors.affiliation: Research Center for Genetic Medicine, Children's National Medical Center, Washington DC, USA.|Department of Integrative Systems Biology, The George Washington University School of Medicine and Health Sciences, Washington, DC, USA.
authors.forename: Jaya
authors.initials: J
authors.lastname: Punetha
authors.affiliation: Research Center for Genetic Medicine, Children's National Medical Center, Washington DC, USA.
authors.forename: Akanchha
authors.initials: A
authors.lastname: Kesari
authors.affiliation: Research Center for Genetic Medicine, Children's National Medical Center, Washington DC, USA.|Department of Integrative Systems Biology, The George Washington University School of Medicine and Health Sciences, Washington, DC, USA.
authors.forename: Prech
authors.initials: P
authors.lastname: Uapinyoying
authors.affiliation: Research Center for Genetic Medicine, Children's National Medical Center, Washington DC, USA.
authors.forename: Mamta
authors.initials: M
authors.lastname: Giri
authors.affiliation: INMR, The Children's Hospital at Westmead & Discipline of Paediatrics and Child Health, University of Sydney, Sydney, Australia.
authors.forename: Nigel F
authors.initials: NF
authors.lastname: Clarke
authors.affiliation: INMR, The Children's Hospital at Westmead & Discipline of Paediatrics and Child Health, University of Sydney, Sydney, Australia.
authors.forename: Leigh B
authors.initials: LB
authors.lastname: Waddell
authors.affiliation: INMR, The Children's Hospital at Westmead & Discipline of Paediatrics and Child Health, University of Sydney, Sydney, Australia.|Murdoch Childrens Research Institute, Melbourne, Australia; Department of Paediatrics, Faculty of Medicine, University of Melbourne, Melbourne, Australia.
authors.forename: Kathryn N
authors.initials: KN
authors.lastname: North
authors.affiliation: INMR, The Children's Hospital at Westmead & Discipline of Paediatrics and Child Health, University of Sydney, Sydney, Australia.
authors.forename: Roula
authors.initials: R
authors.lastname: Ghaoui
authors.affiliation: INMR, The Children's Hospital at Westmead & Discipline of Paediatrics and Child Health, University of Sydney, Sydney, Australia.
authors.forename: Gina L
authors.initials: GL
authors.lastname: O'Grady
authors.affiliation: INMR, The Children's Hospital at Westmead & Discipline of Paediatrics and Child Health, University of Sydney, Sydney, Australia.
authors.forename: Emily C
authors.initials: EC
authors.lastname: Oates
authors.affiliation: INMR, The Children's Hospital at Westmead & Discipline of Paediatrics and Child Health, University of Sydney, Sydney, Australia.
authors.forename: Sarah A
authors.initials: SA
authors.lastname: Sandaradura
authors.affiliation: National Institute of Neurological Disorders and Stroke/NIH, Porter Neuroscience Research Center, Bethesda, MD, USA.
authors.forename: Carsten G
authors.initials: CG
authors.lastname: Bönnemann
authors.affiliation: National Institute of Neurological Disorders and Stroke/NIH, Porter Neuroscience Research Center, Bethesda, MD, USA.
authors.forename: Sandra
authors.initials: S
authors.lastname: Donkervoort
authors.affiliation: National Institute of Arthritis and Musculoskeletal and Skin Diseases, National Institutes of Health, Bethesda, MD, USA.
authors.forename: Paul H
authors.initials: PH
authors.lastname: Plotz
authors.affiliation: Department of Pediatrics, Division of Pediatric Neurology, Duke University Medical Center, Durham, NC, USA.
authors.forename: Edward C
authors.initials: EC
authors.lastname: Smith
authors.affiliation: Research Center for Genetic Medicine, Children's National Medical Center, Washington DC, USA.
authors.forename: Carolina
authors.initials: C
authors.lastname: Tesi-Rocha
authors.affiliation: Department of Neurology, The University of Tennessee Health Science Center, Memphis, TN, USA.
authors.forename: Tulio E
authors.initials: TE
authors.lastname: Bertorini
authors.affiliation: Departments of Pediatrics and Medicine, McMaster University, Neuromuscular Disease Clinic, Health Sciences Centre, ON, Canada.
authors.forename: Mark A
authors.initials: MA
authors.lastname: Tarnopolsky
authors.affiliation: Children's Hospital, Johannes Gutenberg University, Mainz, Germany.
authors.forename: Bernd
authors.initials: B
authors.lastname: Reitter
authors.affiliation: Neuromuscular Unit, Mossakowski Medical Research Centre, Polish Academy of Sciences, Warsaw, Poland.
authors.forename: Irena
authors.initials: I
authors.lastname: Hausmanowa-Petrusewicz
authors.affiliation: Research Center for Genetic Medicine, Children's National Medical Center, Washington DC, USA.|Department of Integrative Systems Biology, The George Washington University School of Medicine and Health Sciences, Washington, DC, USA.
authors.forename: Eric P
authors.initials: EP
authors.lastname: Hoffman
mesh_terms: D005820:Genetic Testing; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D025202:Molecular Diagnostic Techniques; D009135:Muscular Diseases / Q000175:diagnosis* / Q000235:genetics; D009136:Muscular Dystrophies / Q000175:diagnosis / Q000235:genetics; D009154:Mutation; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA
keywords: Exome; Muscular dystrophy; genetic screening; high-throughput DNA sequencing; massively-parallel sequencing; molecular diagnostic testing; molecular diagnostics; myopathies