title: Targeted next-generation sequencing as a comprehensive test for patients with and female carriers of DMD/BMD: a multi-population diagnostic study.
abstract: Duchenne and Becker muscular dystrophies (DMD/BMD) are the most commonly inherited neuromuscular disease. However, accurate and convenient molecular diagnosis cannot be achieved easily because of the enormous size of the dystrophin gene and complex causative mutation spectrum. Such traditional methods as multiplex ligation-dependent probe amplification plus Sanger sequencing require multiple steps to fulfill the diagnosis of DMD/BMD. Here, we introduce a new single-step method for the genetic analysis of DMD patients and female carriers in real clinical settings and demonstrate the validation of its accuracy. A total of 89 patients, 18 female carriers and 245 non-DMD patients were evaluated using our targeted NGS approaches. Compared with traditional methods, our new method yielded 99.99% specificity and 98.96% sensitivity for copy number variations detection and 100% accuracy for the identification of single-nucleotide variation mutations. Additionally, this method is able to detect partial deletions/duplications, thus offering precise personal DMD gene information for gene therapy. We detected novel partial deletions of exons in nine samples for which the breakpoints were located within exonic regions. The results proved that our new method is suitable for routine clinical practice, with shorter turnaround time, higher accuracy, and better insight into comprehensive genetic information (detailed breakpoints) for ensuing gene therapy.
authors.affiliation: BGI-Shenzhen, Shenzhen, China.
authors.forename: Xiaoming
authors.initials: X
authors.lastname: Wei
authors.affiliation: Department of Neurology, Peking Union Medical College Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Dai
authors.affiliation: Department of Medical Genetics School of Medicine, Zhejiang University, Hangzhou, China.
authors.forename: Ping
authors.initials: P
authors.lastname: Yu
authors.affiliation: BGI-Shenzhen, Shenzhen, China.
authors.forename: Ning
authors.initials: N
authors.lastname: Qu
authors.affiliation: BGI-Shenzhen, Shenzhen, China.
authors.forename: Zhangzhang
authors.initials: Z
authors.lastname: Lan
authors.affiliation: Peking Union Medical College, Beijing, China.
authors.forename: Xiafei
authors.initials: X
authors.lastname: Hong
authors.affiliation: BGI-Shenzhen, Shenzhen, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Sun
authors.affiliation: BGI-Shenzhen, Shenzhen, China.
authors.forename: Guanghui
authors.initials: G
authors.lastname: Yang
authors.affiliation: BGI-Shenzhen, Shenzhen, China.
authors.forename: Shuqi
authors.initials: S
authors.lastname: Xie
authors.affiliation: BGI-Shenzhen, Shenzhen, China.
authors.forename: Quan
authors.initials: Q
authors.lastname: Shi
authors.affiliation: BGI-Shenzhen, Shenzhen, China.
authors.forename: Hanlin
authors.initials: H
authors.lastname: Zhou
authors.affiliation: BGI-Shenzhen, Shenzhen, China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Zhu
authors.affiliation: BGI-Shenzhen, Shenzhen, China.
authors.forename: Yuxing
authors.initials: Y
authors.lastname: Chu
authors.affiliation: Clinical Research Lab, Peking Union Medical College Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, China.
authors.forename: Fengxia
authors.initials: F
authors.lastname: Yao
authors.affiliation: BGI-Shenzhen, Shenzhen, China.
authors.forename: Jinming
authors.initials: J
authors.lastname: Wang
authors.affiliation: XiangYa School of Medicine, Central South University, Changsha, China.
authors.forename: Jingni
authors.initials: J
authors.lastname: He
authors.affiliation: BGI-Shenzhen, Shenzhen, China.
authors.forename: Yun
authors.initials: Y
authors.lastname: Yang
authors.affiliation: BGI-Shenzhen, Shenzhen, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Liang
authors.affiliation: BGI-Shenzhen, Shenzhen, China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Yang
authors.affiliation: 1] BGI-Shenzhen, Shenzhen, China [2] Center for Genetic and Genomic Medicine, Zhejiang University School of Medicine First Affiliated Hospital and James D. Watson Institute of Genome Sciences, Hangzhou, China [3] Department of Pathology, University of Rochester Medical Center, Rochester, NY, USA.
authors.forename: Ming
authors.initials: M
authors.lastname: Qi
authors.affiliation: BGI-Tianjin, Tianjin, China.
authors.forename: Ling
authors.initials: L
authors.lastname: Yang
authors.affiliation: BGI-Shenzhen, Shenzhen, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Wang
authors.affiliation: BGI-Shenzhen, Shenzhen, China.
authors.forename: Haitao
authors.initials: H
authors.lastname: Wu
authors.affiliation: Institute of Basic Medical Sciences and School of Basic Medicine, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Duan
authors.affiliation: Peking Union Medical College, Beijing, China.
authors.forename: Cheng
authors.initials: C
authors.lastname: Shen
authors.affiliation: 1] BGI-Shenzhen, Shenzhen, China [2] Department of Biology, University of Copenhagen, Copenhagen, Denmark [3] King Abdulaziz University, Jeddah, Saudi Arabia [4] The Novo Nordisk Foundation Center for Basic Metabolic Research, University of Copenhagen, Copenhagen, Denmark [5] Centre for iSequencing, Aarhus University, Aarhus C, Denmark.
authors.forename: Jun
authors.initials: J
authors.lastname: Wang
authors.affiliation: Department of Neurology, Peking Union Medical College Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, China.
authors.forename: Liying
authors.initials: L
authors.lastname: Cui
authors.affiliation: 1] BGI-Shenzhen, Shenzhen, China [2] BGI-Tianjin, Tianjin, China.
authors.forename: Xin
authors.initials: X
authors.lastname: Yi
mesh_terms: D000328:Adult; D002675:Child, Preschool; D056915:DNA Copy Number Variations / Q000235:genetics; D005260:Female; D006580:Genetic Carrier Screening; D005828:Genetics, Population; D064368:Healthy Volunteers; D006579:Heterozygote; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D006801:Humans; D008297:Male; D008875:Middle Aged; D020388:Muscular Dystrophy, Duchenne / Q000175:diagnosis* / Q000235:genetics* / Q000473:pathology; D057089:Pathology, Molecular; D017384:Sequence Deletion