A Reliable Targeted Next-Generation Sequencing Strategy for Diagnosis of Myopathies and Muscular Dystrophies, Especially for the Giant Titin and Nebulin Genes.
The Journal of Molecular Diagnostics. 2018-07-01; 20(4): 533-549
DOI: 10.1016/j.jmoldx.2018.04.001
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1. J Mol Diagn. 2018 Jul;20(4):533-549. doi: 10.1016/j.jmoldx.2018.04.001. Epub 2018
May 21.
A Reliable Targeted Next-Generation Sequencing Strategy for Diagnosis of
Myopathies and Muscular Dystrophies, Especially for the Giant Titin and Nebulin
Genes.
Zenagui R(1), Lacourt D(1), Pegeot H(1), Yauy K(1), Juntas Morales R(2), Theze
C(1), Rivier F(3), Cances C(4), Sole G(5), Renard D(6), Walther-Louvier U(7),
Ferrer-Monasterio X(5), Espil C(8), Arné-Bes MC(9), Cintas P(9), Uro-Coste E(10),
Martin Negrier ML(11), Rigau V(12), Bieth E(13), Goizet C(14), Claustres M(15),
Koenig M(16), Cossée M(17).
Author information:
(1)Molecular Diagnostic Laboratory, Centre Hospitalier Universitaire Montpellier,
Montpellier, France.
(2)Department of Neurology, Centre Hospitalier Universitaire Montpellier,
Montpellier, France; AOC (Atlantique-Occitanie-Caraïbe) Reference Center for
Neuromuscular Disorders, Aquitaine, France.
(3)AOC (Atlantique-Occitanie-Caraïbe) Reference Center for Neuromuscular
Disorders, Aquitaine, France; Department of Neuropaediatrics, Centre Hospitalier
Universitaire Montpellier, Montpellier, France; PhyMedExp, INSERM, CNRS,
Université de Montpellier, Montpellier, France.
(4)AOC (Atlantique-Occitanie-Caraïbe) Reference Center for Neuromuscular
Disorders, Aquitaine, France; Department of Neuropaediatrics, Centre Hospitalier
Universitaire, Toulouse, France.
(5)AOC (Atlantique-Occitanie-Caraïbe) Reference Center for Neuromuscular
Disorders, Aquitaine, France; Service of Neurology, Centre Hospitalier
Universitaire, Bordeaux, France.
(6)AOC (Atlantique-Occitanie-Caraïbe) Reference Center for Neuromuscular
Disorders, Aquitaine, France; Department of Neurology, Centre Hospitalier
Universitaire, Nîmes, France.
(7)AOC (Atlantique-Occitanie-Caraïbe) Reference Center for Neuromuscular
Disorders, Aquitaine, France; Department of Neuropaediatrics, Centre Hospitalier
Universitaire Montpellier, Montpellier, France.
(8)AOC (Atlantique-Occitanie-Caraïbe) Reference Center for Neuromuscular
Disorders, Aquitaine, France; Department of Neuropaediatrics, Centre Hospitalier
Universitaire, Bordeaux, France.
(9)AOC (Atlantique-Occitanie-Caraïbe) Reference Center for Neuromuscular
Disorders, Aquitaine, France; Department of Neurology, Centre Hospitalier
Universitaire, Toulouse, France.
(10)AOC (Atlantique-Occitanie-Caraïbe) Reference Center for Neuromuscular
Disorders, Aquitaine, France; Department of Pathology, Centre Hospitalier
Universitaire, Toulouse, France.
(11)AOC (Atlantique-Occitanie-Caraïbe) Reference Center for Neuromuscular
Disorders, Aquitaine, France; Service of Pathology, Centre Hospitalier
Universitaire, Bordeaux, France.
(12)AOC (Atlantique-Occitanie-Caraïbe) Reference Center for Neuromuscular
Disorders, Aquitaine, France; Department of Pathology, Centre Hospitalier
Universitaire Montpellier, Montpellier, France.
(13)AOC (Atlantique-Occitanie-Caraïbe) Reference Center for Neuromuscular
Disorders, Aquitaine, France; Department of Genetics, Centre Hospitalier
Universitaire, Toulouse, France.
(14)AOC (Atlantique-Occitanie-Caraïbe) Reference Center for Neuromuscular
Disorders, Aquitaine, France; Department of Genetics, Centre Hospitalier de
Bordeaux, France.
(15)Rare Diseases Genetics Laboratory, Equipe Accueil EA7402, Université de
Montpellier, Montpellier, France.
(16)Molecular Diagnostic Laboratory, Centre Hospitalier Universitaire
Montpellier, Montpellier, France; Rare Diseases Genetics Laboratory, Equipe
Accueil EA7402, Université de Montpellier, Montpellier, France.
(17)Molecular Diagnostic Laboratory, Centre Hospitalier Universitaire
Montpellier, Montpellier, France; AOC (Atlantique-Occitanie-Caraïbe) Reference
Center for Neuromuscular Disorders, Aquitaine, France; Rare Diseases Genetics
Laboratory, Equipe Accueil EA7402, Université de Montpellier, Montpellier,
France. Electronic address: .
Myopathies and muscular dystrophies (M-MDs) are genetically heterogeneous
diseases, with >100 identified genes, including the giant and complex titin (TTN)
and nebulin (NEB) genes. Next-generation sequencing technology revolutionized
M-MD diagnosis and revealed high frequency of TTN and NEB variants. We developed
a next-generation sequencing diagnostic strategy targeted to the coding sequences
of 135 M-MD genes. Comparison of two targeted capture technologies (SeqCap EZ
Choice library capture kit and Nextera Rapid Capture Custom Enrichment kit) and
of two whole-exome sequencing kits (SureSelect V5 and TruSeq RapidExome capture)
revealed best coverage with the SeqCap EZ Choice protocol. A marked decrease in
coverage was observed with the other kits, affecting mostly the first exons of
genes and the repeated regions of TTN and NEB. Bioinformatics analysis strategy
was fine-tuned to achieve optimal detection of variants, including small
insertions/deletions (INDELs) and copy number variants (CNVs). Analysis of a
cohort of 128 patients allowed the detection of 52 substitutions, 13 INDELs
(including a trinucleotide repeat expansion), and 3 CNVs. Two INDELs were
localized in the repeated regions of NEB, suggesting that these mutations may be
frequent but underestimated. A large deletion was also identified in TTN that is,
to our knowledge, the first published CNV in this gene.
Copyright © 2018 American Society for Investigative Pathology and the Association
for Molecular Pathology. Published by Elsevier Inc. All rights reserved.
DOI: 10.1016/j.jmoldx.2018.04.001
PMID: 29792937 [Indexed for MEDLINE]