title: Targeted sequence capture and GS-FLX Titanium sequencing of 23 hypertrophic and dilated cardiomyopathy genes: implementation into diagnostics.
abstract: BACKGROUND
  Genetic evaluation of cardiomyopathies poses a challenge. Multiple genes are involved but no clear genotype-phenotype correlations have been found so far. In the past, genetic evaluation for hypertrophic (HCM) and dilated (DCM) cardiomyopathies was performed by sequential screening of a very limited number of genes. Recent developments in sequencing have increased the throughput, enabling simultaneous screening of multiple genes for multiple patients in a single sequencing run.
  
  
  OBJECTIVE
  Development and implementation of a next generation sequencing (NGS) based genetic test as replacement for Sanger sequencing.
  
  
  RESULTS
  In order to increase the number of genes that can be screened in a shorter time period, we enriched all exons of 23 of the most relevant HCM and DCM related genes using on-array multiplexed sequence capture followed by massively parallel pyrosequencing on the GS-FLX Titanium. After optimisation of array based sequence capture it was feasible to reliably detect a large panel of known and unknown variants in HCM and DCM patients, whereby the unknown variants could be confirmed by Sanger sequencing.
  
  
  CONCLUSIONS
  The rate of detection of (pathogenic) variants in both HCM and DCM patients was increased due to a larger number of genes studied. Array based target enrichment followed by NGS showed the same accuracy as Sanger sequencing. Therefore, NGS is ready for implementation in a diagnostic setting.
authors.affiliation: Department of Clinical Genetics, Academic Medical Center, Amsterdam, The Netherlands. o.r.mook@amc.uva.nl
authors.forename: Olaf R F
authors.initials: OR
authors.lastname: Mook
authors.forename: Martin A
authors.initials: MA
authors.lastname: Haagmans
authors.forename: Jean-François
authors.initials: JF
authors.lastname: Soucy
authors.forename: Judith B A
authors.initials: JB
authors.lastname: van de Meerakker
authors.forename: Frank
authors.initials: F
authors.lastname: Baas
authors.forename: Marja E
authors.initials: ME
authors.lastname: Jakobs
authors.forename: Nynke
authors.initials: N
authors.lastname: Hofman
authors.forename: Imke
authors.initials: I
authors.lastname: Christiaans
authors.forename: Ronald H
authors.initials: RH
authors.lastname: Lekanne Deprez
authors.forename: Marcel M A M
authors.initials: MM
authors.lastname: Mannens
mesh_terms: D000328:Adult; D000368:Aged; D006332:Cardiomegaly / Q000235:genetics*; D002311:Cardiomyopathy, Dilated / Q000235:genetics; D005260:Female; D005820:Genetic Testing / Q000379:methods*; D006801:Humans; D008297:Male; D008875:Middle Aged; D020641:Polymorphism, Single Nucleotide; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA / Q000379:methods*; D014025:Titanium / Q000737:chemistry*
chemical_list: D014025:Titanium
keywords: Cardiomyopathy; Diagnostics; Genetics; Molecular genetics