TY - JOUR
T1 - Targeted sequence capture and GS-FLX Titanium sequencing of 23 hypertrophic and dilated cardiomyopathy genes: implementation into diagnostics
AU - Mook, Olaf R. F.
AU - Haagmans, Martin A.
AU - Soucy, Jean-François
AU - van de Meerakker, Judith B. A.
AU - Baas, Frank
AU - Jakobs, Marja E.
AU - Hofman, Nynke
AU - Christiaans, Imke
AU - Lekanne Deprez, Ronald H.
AU - Mannens, Marcel M. A. M.
PY - 2013
Y1 - 2013
N2 - 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. Development and implementation of a next generation sequencing (NGS) based genetic test as replacement for Sanger sequencing. 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. 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
AB - 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. Development and implementation of a next generation sequencing (NGS) based genetic test as replacement for Sanger sequencing. 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. 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
U2 - https://doi.org/10.1136/jmedgenet-2012-101231
DO - https://doi.org/10.1136/jmedgenet-2012-101231
M3 - Article
C2 - 23785128
SN - 0022-2593
VL - 50
SP - 614
EP - 626
JO - Journal of medical genetics
JF - Journal of medical genetics
IS - 9
ER -