title: Targeted next-generation sequencing for the genetic diagnosis of dysferlinopathy.
abstract: Dysferlinopathy comprises a group of autosomal recessive muscular dystrophies caused by mutations in the DYSF gene. Due to the large size of the gene and its lack of mutational hot spots, analysis of the DYSF gene is time-consuming and laborious using conventional sequencing methods. By next-generation sequencing (NGS), DYSF gene analysis has previously been validated through its incorporation in multi-gene panels or exome analyses. However, individual validation of NGS approaches for DYSF gene has not been performed. Here, we established and validated a hybridization capture-based target-enrichment followed by next-generation sequencing to detect mutations in patients with dysferlinopathy. With this approach, mean depth of coverage was approximately 450 fold and almost all (99.3%) of the targeted region had sequence coverage greater than 20 fold. When this approach was tested on samples from patients with known DYSF mutations, all known mutations were correctly retrieved. Using this method on 32 consecutive patient samples with dysferlinopathy, at least two pathogenic variants were detected in 28 (87.5%) samples and at least one pathogenic variant was identified in all samples. Our results suggested that the NGS-based screening method could facilitate efficient and accurate genetic diagnosis of dysferlinopathy.
authors: Department of Neurology, Yonsei University College of Medicine, Seoul, Republic of Korea.
authors: Ha Young
authors: HY
authors: Shin
authors: Department of Chemistry, Yonsei University, Seoul, Republic of Korea.
authors: Hoon
authors: H
authors: Jang
authors: Department of Pharmacology, Pharmacogenomic Research Center for Membrane Transporters, Brain Korea 21 PLUS Project for Medical Sciences, Severance Biomedical Science Institute, Yonsei University College of Medicine, Seoul, Republic of Korea.
authors: Joo Hyung
authors: JH
authors: Han
authors: Department of Neurology, Mokdong Hospital, Ewha Womans University College of Medicine, Seoul, Republic of Korea.
authors: Hyung Jun
authors: HJ
authors: Park
authors: Department of Neurology, Yonsei University College of Medicine, Seoul, Republic of Korea.
authors: Jung Hwan
authors: JH
authors: Lee
authors: Department of Pharmacology and PharmacoGenomics Research Center, Inje University College of Medicine, Busan, Republic of Korea; Department of Clinical Pharmacology, Inje University, Busan Paik Hospital, Busan, Republic of Korea.
authors: So Won
authors: SW
authors: Kim
authors: Department of Neurology, Yonsei University College of Medicine, Seoul, Republic of Korea.
authors: Seung Min
authors: SM
authors: Kim
authors: Department of Neurology, Pusan National University School of Medicine, Busan, Republic of Korea.
authors: Young-Eun
authors: YE
authors: Park
authors: Department of Neurology, Pusan National University School of Medicine, Busan, Republic of Korea.
authors: Dae-Seong
authors: DS
authors: Kim
authors: Department of Chemistry, Yonsei University, Seoul, Republic of Korea.
authors: Duhee
authors: D
authors: Bang
authors: Department of Pharmacology, Pharmacogenomic Research Center for Membrane Transporters, Brain Korea 21 PLUS Project for Medical Sciences, Severance Biomedical Science Institute, Yonsei University College of Medicine, Seoul, Republic of Korea.
authors: Min Goo
authors: MG
authors: Lee
authors: Department of Oral Biology, Yonsei University College of Dentistry, Seoul, Republic of Korea. Electronic address: jihyni@yuhs.ac.
authors: Ji Hyun
authors: JH
authors: Lee
authors: Department of Neurology, Yonsei University College of Medicine, Seoul, Republic of Korea. Electronic address: ycchoi@yuhs.ac.
authors: Young-Chul
authors: YC
authors: Choi
mesh_terms: D000293:Adolescent; D000328:Adult; D002648:Child; D004252:DNA Mutational Analysis; D000073939:Dysferlin; D005260:Female; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D008297:Male; D008565:Membrane Proteins / Q000235:genetics*; D009124:Muscle Proteins / Q000235:genetics*; D049288:Muscular Dystrophies, Limb-Girdle / Q000175:diagnosis* / Q000235:genetics*; D055815:Young Adult
chemical_list: C114839:DYSF protein, human; D000073939:Dysferlin; D008565:Membrane Proteins; D009124:Muscle Proteins
keywords: DYSF; Dysferlinopathy; Hybridization capture; Mutation; Next-generation sequencing