Highlights
- •DYSF gene analysis is time-consuming and laborious by conventional sequencing method.
- •We set up a next-generation sequencing-based screening method for dysferlinopathy.
- •This method is accurate and efficient for 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.
Keywords
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Article info
Publication history
Published online: March 17, 2015
Accepted:
March 11,
2015
Received in revised form:
February 16,
2015
Received:
December 4,
2014
Identification
Copyright
© 2015 Elsevier B.V. Published by Elsevier Inc. All rights reserved.
