An Extended Targeted Copy Number Variation Detection Array Including 187 Genes for the Diagnostics of Neuromuscular Disorders

J Neuromuscul Dis. 2018;5(3):307-314. doi: 10.3233/JND-170298.

Abstract

Background: Our previous array, the Comparative Genomic Hybridisation design (CGH-array) for nemaline myopathy (NM), named the NM-CGH array, revealed pathogenic copy number variation (CNV) in the genes for nebulin (NEB) and tropomyosin 3 (TPM3), as well as recurrent CNVs in the segmental duplication (SD), i.e. triplicate, region of NEB (TRI, exons 82-89, 90-97, 98-105). In the light of this knowledge, we have designed and validated an extended CGH array, which includes a selection of 187 genes known to cause neuromuscular disorders (NMDs).

Objective: Our aim was to develop a reliable method for CNV detection in genes related to neuromuscular disorders for routine mutation detection and analysis, as a much-needed complement to sequencing methods.

Methods: We have developed a novel custom-made 4×180 k CGH array for the diagnostics of NMDs. It includes the same tiled ultra-high density coverage of the 12 known or putative NM genes as our 8×60 k NM-CGH-array but also comprises a selection of 175 additional genes associated with NMDs, including titin (TTN), at a high to very high coverage. The genes were divided into three coverage groups according to known and potential pathogenicity in neuromuscular disorders.

Results: The array detected known and putative CNVs in all three gene coverage groups, including the repetitive regions of NEB and TTN.

Conclusions: The targeted neuromuscular disorder 4×180 k array-CGH (NMD-CGH-array v1.0) design allows CNV detection for a broader spectrum of neuromuscular disorders at a high resolution.

Keywords: Array comparative genomic hybridisation (aCGH); NEB; NM-CGH; NMD-CGH; TTN; copy number variation (CNV); nebulin; titin.

MeSH terms

  • Adult
  • Connectin / genetics
  • DNA / genetics
  • DNA Copy Number Variations / genetics*
  • Female
  • Genomics / methods
  • Humans
  • Male
  • Microarray Analysis
  • Muscle Proteins / genetics
  • Mutation / genetics
  • Myopathies, Nemaline / genetics
  • Neuromuscular Diseases / diagnosis*
  • Neuromuscular Diseases / genetics*
  • Oligonucleotide Array Sequence Analysis
  • Reproducibility of Results

Substances

  • Connectin
  • Muscle Proteins
  • TTN protein, human
  • nebulin
  • DNA