Targeted Next-Generation Sequencing of 51 Genes Involved in Primary Electrical Disease

Abstract

Primary electrical disease (PED) is characterized by cardiac arrhythmias, which can lead to sudden cardiac death in the absence of detectable structural heart disease. PED encompasses a diversity of inherited syndromes, predominantly Brugada syndrome, early repolarization syndrome, long QT syndrome, short QT syndrome, arrhythmogenic right ventricular cardiomyopathy, and catecholaminergic polymorphic ventricular tachycardia. To overcome the diagnostic challenges imposed by the clinical and genetic heterogeneity of PED, we developed a targeted gene panel for next-generation sequencing of 51 PED genes. The amplified samples were sequenced on MiSeq. To validate the panel, 20 Human Polymorphism Study Center samples and 19 positive control samples were used, with a total of 1479 variants. An analytical sensitivity and specificity of 100% and 99.9% were obtained. After validation, we applied the assay to 114 PED patients. We identified 107 variants in 36 different genes, 18 of which were classified as pathogenic or likely pathogenic, 54 variants were of unknown significance, and 35 were classified as likely benign. We can conclude that the PED Multiplex Amplification of Specific Targets for Resequencing Plus assay is a proficient and highly reliable test to routinely screen patients experiencing primary arrhythmias.

NKX2-5 Variant in Two Siblings with Thyroid Hemiagenesis

“…Based on these findings, genes expressed during embryogenesis in precursors of thyroid and heart are promising candidate genes to be considered for their involvement in the pathogenesis of TD. The variant detected in our patients’ family has been so far reported to be causative in an individual affected with accessory atrioventricular connection and one individual referred for primary electrical disease testing [ 9 , 10 ]. However, neither this variant nor any other mutation in NKX2-5 gene were so far detected in patients with THA.…”

Molecular autopsy and subsequent functional analysis reveal de novo DSG2 mutation as cause of sudden death

“…After inverse centrifuging the serum tubes, we were able to recover white blood cells and obtained sufficient quality and amount of DNA. After informed consent and permission of his spouse post-mortem genetic analysis was performed on this sample using next-generation sequencing panels of known cardiomyopathy and primary arrhythmia genes (each n=51 genes) (13). Segregation analysis showed that the KCNQ1 variant was inherited from the proband's mother and transmitted to the proband's daughter.…”

Compound Heterozygous SCN5A Mutations in Severe Sodium Channelopathy With Brugada Syndrome: A Case Report

“…Genomic DNA was extracted from whole blood using standard procedures. Genetic testing of SCN5A was performed using an in-house developed cardiac arrhythmia gene panel (14). Sanger sequencing was used to validate the variants and perform familial segregation analysis.…”