Development of a Comprehensive Sequencing Assay for Inherited Cardiac Condition Genes

J Cardiovasc Transl Res. 2016 Feb;9(1):3-11. doi: 10.1007/s12265-016-9673-5. Epub 2016 Feb 17.

Abstract

Inherited cardiac conditions (ICCs) are characterised by marked genetic and allelic heterogeneity and require extensive sequencing for genetic characterisation. We iteratively optimised a targeted gene capture panel for ICCs that includes disease-causing, putatively pathogenic, research and phenocopy genes (n = 174 genes). We achieved high coverage of the target region on both MiSeq (>99.8% at ≥ 20× read depth, n = 12) and NextSeq (>99.9% at ≥ 20×, n = 48) platforms with 100% sensitivity and precision for single nucleotide variants and indels across the protein-coding target on the MiSeq. In the final assay, 40 out of 43 established ICC genes informative in clinical practice achieved complete coverage (100 % at ≥ 20×). By comparison, whole exome sequencing (WES; ∼ 80×), deep WES (∼ 500×) and whole genome sequencing (WGS; ∼ 70×) had poorer performance (88.1, 99.2 and 99.3% respectively at ≥ 20×) across the ICC target. The assay described here delivers highly accurate and affordable sequencing of ICC genes, complemented by accessible cloud-based computation and informatics. See Editorial in this issue (DOI: 10.1007/s12265-015-9667-8 ).

Keywords: Diagnostics; Genetics; Inherited cardiac conditions; Targeted sequencing; Whole exome sequencing; Whole genome sequencing.

Publication types

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

MeSH terms

  • Cloud Computing
  • Computational Biology
  • DNA Mutational Analysis / methods*
  • Databases, Genetic
  • Exome
  • Genetic Markers
  • Genetic Predisposition to Disease
  • Heart Diseases / diagnosis
  • Heart Diseases / genetics*
  • Heredity
  • High-Throughput Nucleotide Sequencing / methods*
  • Humans
  • London
  • Mutation*
  • Phenotype
  • Polymorphism, Single Nucleotide*
  • Predictive Value of Tests
  • Singapore
  • Workflow

Substances

  • Genetic Markers