[HTML][HTML] Development of a comprehensive sequencing assay for inherited cardiac condition genes

CJ Pua, J Bhalshankar, K Miao, R Walsh… - Journal of …, 2016 - Springer
CJ Pua, J Bhalshankar, K Miao, R Walsh, S John, SQ Lim, K Chow, R Buchan, BY Soh…
Journal of cardiovascular translational research, 2016Springer
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 …
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 ).
Springer