RT Journal Article
SR Electronic
T1 Expanding genotype/phenotype of neuromuscular diseases by comprehensive target capture/NGS
JF Neurology Genetics
JO Neurol Genet
FD Lippincott Williams & Wilkins
SP e14
DO 10.1212/NXG.0000000000000015
VO 1
IS 2
A1 Xia Tian
A1 Wen-Chen Liang
A1 Yanming Feng
A1 Jing Wang
A1 Victor Wei Zhang
A1 Chih-Hung Chou
A1 Hsien-Da Huang
A1 Ching Wan Lam
A1 Ya-Yun Hsu
A1 Thy-Sheng Lin
A1 Wan-Tzu Chen
A1 Lee-Jun Wong
A1 Yuh-Jyh Jong
YR 2015
UL http://ng.neurology.org/content/1/2/e14.abstract
AB Objective: To establish and evaluate the effectiveness of a comprehensive next-generation sequencing (NGS) approach to simultaneously analyze all genes known to be responsible for the most clinically and genetically heterogeneous neuromuscular diseases (NMDs) involving spinal motoneurons, neuromuscular junctions, nerves, and muscles.Methods: All coding exons and at least 20 bp of flanking intronic sequences of 236 genes causing NMDs were enriched by using SeqCap EZ solution-based capture and enrichment method followed by massively parallel sequencing on Illumina HiSeq2000.Results: The target gene capture/deep sequencing provides an average coverage of ∼1,000× per nucleotide. Thirty-five unrelated NMD families (38 patients) with clinical and/or muscle pathologic diagnoses but without identified causative genetic defects were analyzed. Deleterious mutations were found in 29 families (83%). Definitive causative mutations were identified in 21 families (60%) and likely diagnoses were established in 8 families (23%). Six families were left without diagnosis due to uncertainty in phenotype/genotype correlation and/or unidentified causative genes. Using this comprehensive panel, we not only identified mutations in expected genes but also expanded phenotype/genotype among different subcategories of NMDs.Conclusions: Target gene capture/deep sequencing approach can greatly improve the genetic diagnosis of NMDs. This study demonstrated the power of NGS in confirming and expanding clinical phenotypes/genotypes of the extremely heterogeneous NMDs. Confirmed molecular diagnoses of NMDs can assist in genetic counseling and carrier detection as well as guide therapeutic options for treatable disorders.AD=autosomal dominant; AR=autosomal recessive; BCM=Baylor College of Medicine; CACTD=carnitine acylcarnitine translocase deficiency; CCD=central core disease; CM=congenital myopathy; CMD=congenital muscular dystrophy; CMT=Charcot-Marie-Tooth disease; CMyS=congenital myasthenic syndrome; CNM=centronuclear myopathy; EDMD=Emery-Dreifuss muscular dystrophy; HSAN=hereditary sensory and autonomic neuropathy; LGMD=limb-girdle muscular dystrophy; MH=malignant hyperthermia; MM=metabolic myopathy; MTS=myotonic syndrome; NGS=next-generation sequencing; NM=nemaline myopathy; NMD=neuromuscular disease; PFIS=paralytic floppy infant syndrome; SMARD1=spinal muscular atrophy with respiratory distress type 1; VUS=variants of unknown significance