Mutation Analysis Ion Channel Genes Ventricular Fibrillation Survivors with Coronary Artery Disease

Novotný, Tomáš; Kadlecová, Jitka; Raudenská, Martina; Bittnerová, Alexandra; Andršová, Irena; Floriánová, Alena; Vašků, Anna; Neugebauer, Petr; Kozák, Milan; Sepši, Milan; Křivan, Lubomír; Gaillyová, Renata; Špinar, Jindřich

doi:10.1111/j.1540-8159.2011.03045.x

Cited by 10 publications

(12 citation statements)

References 23 publications

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“…Fifteen anonymised DNA samples were selected for technical assay evaluation. Eleven (group I) had undergone mutation scanning of five Long QT syndrome (LQT) associated genes (See Table 1) using denaturing high performance liquid chromatography (dHPLC) [21] coupled with Sanger DNA sequence analysis to confirm putative variants. Four (group II) underwent exon PCR amplification and direct Sanger DNA sequence analysis of the full coding sequence of the same five genes.…”

Section: Methodsmentioning

confidence: 99%

Towards Clinical Molecular Diagnosis of Inherited Cardiac Conditions: A Comparison of Bench-Top Genome DNA Sequencers

Buckton

Wilkinson

et al. 2013

PLoS ONE

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BackgroundMolecular genetic testing is recommended for diagnosis of inherited cardiac disease, to guide prognosis and treatment, but access is often limited by cost and availability. Recently introduced high-throughput bench-top DNA sequencing platforms have the potential to overcome these limitations.Methodology/Principal FindingsWe evaluated two next-generation sequencing (NGS) platforms for molecular diagnostics. The protein-coding regions of six genes associated with inherited arrhythmia syndromes were amplified from 15 human samples using parallelised multiplex PCR (Access Array, Fluidigm), and sequenced on the MiSeq (Illumina) and Ion Torrent PGM (Life Technologies). Overall, 97.9% of the target was sequenced adequately for variant calling on the MiSeq, and 96.8% on the Ion Torrent PGM. Regions missed tended to be of high GC-content, and most were problematic for both platforms. Variant calling was assessed using 107 variants detected using Sanger sequencing: within adequately sequenced regions, variant calling on both platforms was highly accurate (Sensitivity: MiSeq 100%, PGM 99.1%. Positive predictive value: MiSeq 95.9%, PGM 95.5%). At the time of the study the Ion Torrent PGM had a lower capital cost and individual runs were cheaper and faster. The MiSeq had a higher capacity (requiring fewer runs), with reduced hands-on time and simpler laboratory workflows. Both provide significant cost and time savings over conventional methods, even allowing for adjunct Sanger sequencing to validate findings and sequence exons missed by NGS.Conclusions/SignificanceMiSeq and Ion Torrent PGM both provide accurate variant detection as part of a PCR-based molecular diagnostic workflow, and provide alternative platforms for molecular diagnosis of inherited cardiac conditions. Though there were performance differences at this throughput, platforms differed primarily in terms of cost, scalability, protocol stability and ease of use. Compared with current molecular genetic diagnostic tests for inherited cardiac arrhythmias, these NGS approaches are faster, less expensive, and yet more comprehensive.

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Section: Methodsmentioning

confidence: 99%

Towards Clinical Molecular Diagnosis of Inherited Cardiac Conditions: A Comparison of Bench-Top Genome DNA Sequencers

Buckton

Wilkinson

et al. 2013

PLoS ONE

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“…VF occurs in 10% of cases within the first hours following the symptoms of an AMI (2,3). In addition to established risk factors for VF, a significant genetic component may be detected using extensive observational population studies (4)(5)(6)(7). In primary electrical heart diseases, including Long QT syndrome (LQTS), Short QT syndrome and Brugada syndrome, genes encoding ion channels are mutated.…”

Section: Introductionmentioning

confidence: 99%

SCN5A mutations and polymorphisms in patients with ventricular fibrillation during acute myocardial infarction

Boehringer

Bugert

Borggrefe

et al. 2014

Molecular Medicine Reports

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Mutations in the SCN5A gene encoding the Nav1.5 channel α-subunit are known to be risk factors of arrhythmia, including Brugada Syndrome and Long QT syndrome subtype 3. The present study focused on the role of SCN5A variants in the development of ventricular fibrillation (VF) during acute myocardial infarction (AMI). Since VF during AMI is the major cause of sudden death in the Western world, SCN5A mutations represent genetic risk factors for sudden death. By exon re-sequencing, the entire coding region and flanking intron regions were sequenced in 46 AMI/VF+ patients. In total, nine single nucleotide variants were identified of which four represented common single nucleotide polymorphisms (SNPs; 87G>A, 1673A>G, IVS16‑6C>T and 5457T>A). Only five rare variants were identified, each in only one patient. Only two of the rare variants represented missense mutations (3578G>A and 4786T>A). The common SNPs and the missense mutations were also genotyped using polymerase chain reaction methods in 79 AMI/VF‑ patients and 480 healthy controls. The SNPs did not demonstrate significant differences in allele and genotype frequencies between the study groups. The 3578G>A mutation was identified in one out of the 480 controls, whereas the 4786T>A mutation was not present in AMI/VF- patients and controls. In conclusion, the majority of AMI/VF+ patients demonstrated a wild type sequence or common SNPs in SCN5A. Only two out of 46 (4.3%) AMI/VF+ patients revealed mutations that may be involved in Nav1.5 dysfunction and VF. However, this requires further functional validation.

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“…[ 19 , 23 , 27 ] Four articles pertained to survivors of SCD who were successfully resuscitated. [ 23 , 25 , 27 , 35 ] The common ion channel-related genes that were assessed in the studies included SCN5A, SCN10A, KCNQ1, KCNH2, KCNE1, and RYR2. The NOS scores of the included studies ranged from 6 to 8.…”

Section: Resultsmentioning

confidence: 99%

Associations between common ion channel single nucleotide polymorphisms and sudden cardiac death in adults

Liu

Shi²,

Xiao³

2018

Medicine

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Mutation Analysis Ion Channel Genes Ventricular Fibrillation Survivors with Coronary Artery Disease

Abstract: The prevalence of selected, rare coding variants in five long QT genes was significantly higher in cases versus controls, confirming a mechanistic role for these genes among a subgroup of patients with coronary disease and VF.

Cited by 10 publications

References 23 publications

Towards Clinical Molecular Diagnosis of Inherited Cardiac Conditions: A Comparison of Bench-Top Genome DNA Sequencers

Towards Clinical Molecular Diagnosis of Inherited Cardiac Conditions: A Comparison of Bench-Top Genome DNA Sequencers

SCN5A mutations and polymorphisms in patients with ventricular fibrillation during acute myocardial infarction

Associations between common ion channel single nucleotide polymorphisms and sudden cardiac death in adults

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