Diagnostic whole genome sequencing and split-read mapping for nucleotide resolution breakpoint identification in CNTNAP2 deficiency syndrome

Am J Med Genet A. 2014 Oct;164A(10):2649-55. doi: 10.1002/ajmg.a.36679. Epub 2014 Jul 16.

Abstract

Whole genome sequencing (WGS) has the potential to report on all types of genetic abnormality, thus converging diagnostic testing on a single methodology. Although WGS at sufficient depth for robust detection of point mutations is still some way from being affordable for diagnostic purposes, low-coverage WGS is already an excellent method for detecting copy number variants ("CNVseq"). We report on a family in which individuals presented with a presumed autosomal recessive syndrome of severe intellectual disability and epilepsy. Array comparative genomic hybridization (CGH) analysis had revealed a homozygous deletion apparently lying within intron 3 of CNTNAP2. Since this was too small for confirmation by FISH, CNVseq was used, refining the extent of this mutation to approximately 76.8 kb, encompassing CNTNAP2 exon 3 (an out-of-frame deletion). To characterize the precise breakpoints and provide a rapid molecular diagnostic test, we resequenced the CNVseq library at medium coverage and performed split read mapping. This yielded information for a multiplex polymerase chain reaction (PCR) assay, used for cascade screening and/or prenatal diagnosis in this family. This example demonstrates a rapid, low-cost approach to converting molecular cytogenetic findings into robust PCR-based tests.

Keywords: CNTNAP2; split read mapping; whole genome sequencing.

Publication types

  • Case Reports

MeSH terms

  • Adolescent
  • DNA Copy Number Variations / genetics
  • Exons / genetics
  • Female
  • Humans
  • Male
  • Membrane Proteins / deficiency*
  • Membrane Proteins / genetics*
  • Molecular Diagnostic Techniques / methods
  • Mutation / genetics
  • Nerve Tissue Proteins / deficiency*
  • Nerve Tissue Proteins / genetics*
  • Nucleotides / genetics*
  • Pedigree
  • Sequence Analysis, DNA / methods
  • Sequence Deletion / genetics*

Substances

  • CNTNAP2 protein, human
  • Membrane Proteins
  • Nerve Tissue Proteins
  • Nucleotides