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Whole dystrophin gene analysis by next-generation sequencing: a comprehensive genetic diagnosis of Duchenne and Becker muscular dystrophy

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Abstract

Duchenne/Becker muscular dystrophies are the most frequent inherited neuromuscular diseases caused by mutations of the dystrophin gene. However, approximately 30 % of patients with the disease do not receive a molecular diagnosis because of the complex mutational spectrum and the large size of the gene. The introduction and use of next-generation sequencing have advanced clinical genetic research and might be a suitable method for the detection of various types of mutations in the dystrophin gene. To identify the mutational spectrum using a single platform, whole dystrophin gene sequencing was performed using next-generation sequencing. The entire dystrophin gene, including all exons, introns and promoter regions, was target enriched using a DMD whole gene enrichment kit. The enrichment libraries were sequenced on an Illumina HiSeq 2000 sequencer using paired read 100 bp sequencing. We studied 26 patients: 21 had known large deletion/duplications and 5 did not have detectable large deletion/duplications by multiplex ligation-dependent probe amplification technology (MLPA). We applied whole dystrophin gene analysis by next-generation sequencing to the five patients who did not have detectable large deletion/duplications and to five randomly chosen patients from the 21 who did have large deletion/duplications. The sequencing data covered almost 100 % of the exonic region of the dystrophin gene by ≥10 reads with a mean read depth of 147. Five small mutations were identified in the first five patients, of which four variants were unreported in the dmd.nl database. The deleted or duplicated exons and the breakpoints in the five large deletion/duplication patients were precisely identified. Whole dystrophin gene sequencing by next-generation sequencing may be a useful tool for the genetic diagnosis of Duchenne and Becker muscular dystrophies.

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Acknowledgments

We thank all the tested individuals, their families and collaborating clinicians for their participation. This work was supported by the National Natural Science Foundation of China (81300527), the National Science and Technology Support Program of China (2013BA112B00) and Beijing Medical Capital Development Foundation 2012. This study has neither been presented nor submitted or accepted anywhere.

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The authors declare no conflict of interest.

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Authors and Affiliations

  1. Department of Clinical Genetics, BaYi Children’s Hospital of the General Military Hospital of Beijing PLA, 5 Nanmencang Road, Dongcheng District, Beijing, 100700, People’ Republic of China

    Yan Wang, Yao Yang, Xiao-Chun Chen, Xin Liu, Chun-Zhi Wang & Xi-Yu He

  2. The Pharmacology Department, General Military Hospital of Beijing PLA, Beijing, 100700, People’ Republic of China

    Jing Liu

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  1. Yan Wang
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  2. Yao Yang
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  3. Jing Liu
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  4. Xiao-Chun Chen
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  5. Xin Liu
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  6. Chun-Zhi Wang
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  7. Xi-Yu He
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Correspondence to Xi-Yu He.

Additional information

Communicated by S. Hohmann.

Y. Wang, Y. Yang, and J. Liu contributed equally to the paper.

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Wang, Y., Yang, Y., Liu, J. et al. Whole dystrophin gene analysis by next-generation sequencing: a comprehensive genetic diagnosis of Duchenne and Becker muscular dystrophy. Mol Genet Genomics 289, 1013–1021 (2014). https://doi.org/10.1007/s00438-014-0847-z

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