Validation of next-generation sequencing technologies in genetic diagnosis of dementia

Neurobiol Aging. 2014 Jan;35(1):261-5. doi: 10.1016/j.neurobiolaging.2013.07.017. Epub 2013 Aug 31.

Abstract

Identification of a specific genetic cause of early onset dementia (EOD) is important but can be difficult because of pleiotropy, locus heterogeneity and accessibility of gene tests. Here we assess the use of next generation sequencing (NGS) technologies as a quick, accurate and cost effective method to determine genetic diagnosis in EOD. We developed gene panel based technologies to assess 16 genes known to harbour mutations causal of dementia and combined these with PCR based assessments of the C9orf72 hexanucleotide repeat expansion and the octapeptide repeat region of PRNP. In a blinded study of 95 samples we show very high sensitivity and specificity are achievable using either Ion Torrent or MiSeq sequencing platforms. Modifications to the gene panel permit accurate detection of structural variation in APP. In 2/10 samples which had been selected because they possess a variant of uncertain significance the new technology discovered a causal mutation in genes not previously sequenced. A large proportion (23/85) of samples showed genetic variants of uncertain significance in addition to known mutations. The MRC Dementia Gene Panel and similar technologies are likely to be transformational in EOD diagnosis with a significant impact on the proportion of patients in whom a genetic cause is identified.

Keywords: Dementia; Diagnosis; Genetic; Ion torrent; MiSeq; NGS; Neurogenetics; Next-generation sequencing; Sequencing.

Publication types

  • Research Support, Non-U.S. Gov't
  • Validation Study

MeSH terms

  • Amyloid beta-Protein Precursor / genetics
  • C9orf72 Protein
  • DNA Repeat Expansion
  • Dementia / diagnosis*
  • Dementia / genetics*
  • Genetic Variation
  • Humans
  • Molecular Diagnostic Techniques / methods*
  • Mutation
  • Polymerase Chain Reaction / methods
  • Prion Proteins
  • Prions / genetics
  • Proteins / genetics
  • Sensitivity and Specificity
  • Sequence Analysis, DNA / methods*

Substances

  • APP protein, human
  • Amyloid beta-Protein Precursor
  • C9orf72 Protein
  • C9orf72 protein, human
  • PRNP protein, human
  • Prion Proteins
  • Prions
  • Proteins