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Candidate Gene Testing in Clinical Cohort Studies with Multiplexed Genotyping and Mass Spectrometry
Published on: June 21, 2018
Multiplex targeted high-throughput sequencing for Mendelian cardiac disorders
S Fokstuen1, P Makrythanasis, S Nikolaev
1Genetic Medicine, University Hospitals of Geneva, Geneva, Switzerland.
Insights
Multiplex targeted high-throughput sequencing (HTS) offers a fast, cost-effective diagnostic method for genetic heart disorders like hypertrophic cardiomyopathy (HCM) and long QT syndrome (LQTS). This approach efficiently identifies pathogenic variants, improving molecular diagnosis for heterogeneous Mendelian cardiac conditions.
Area of Science:
- Cardiovascular Genetics
- Molecular Diagnostics
- Next-Generation Sequencing
Background:
- Mendelian cardiomyopathies and arrhythmias exhibit significant genetic heterogeneity.
- Traditional Sanger sequencing is laborious and costly for diagnosing these diverse genetic disorders.
Purpose of the Study:
- To evaluate multiplex targeted high-throughput sequencing (HTS) as a rapid and cost-efficient diagnostic approach for Mendelian cardiac disorders.
- To demonstrate the feasibility of simultaneously analyzing multiple patient samples using multiplexing.
Main Methods:
- Designed a DNA capture assay targeting all exons of 130 genes associated with cardiovascular Mendelian disorders.
- Applied multiplex targeted HTS to analyze four patient samples concurrently.
- Investigated two familial hypertrophic cardiomyopathy (HCM) cases and two long QT syndrome (LQTS) cases.
Main Results:
- Identified known pathogenic variants in MYH7 and MYBPC in an HCM patient.
- Detected an acceptor splice site variant in MYPC3 in a second HCM patient.
- Found missense variants in SCN5A and KCNQ in an LQTS patient.
- Confirmed a missense variant in MYBPC3 in an LQTS patient, a gene typically associated with cardiomyopathy.
Conclusions:
- Multiplex targeted HTS is an efficient and cost-effective tool for the molecular diagnosis of genetically heterogeneous cardiac disorders.
- This method provides valuable insights into the pathogenesis of complex Mendelian cardiac diseases.
- Facilitates improved clinical practice for diagnosing inherited heart conditions.
Abstract:
Mendelian cardiomyopathies and arrhythmias are characterized by an important genetic heterogeneity, rendering Sanger sequencing very laborious and expensive. As a proof of concept, we explored multiplex targeted high-throughput sequencing (HTS) as a fast and cost-efficient diagnostic method for individuals suffering from Mendelian cardiac disorders. We designed a DNA capture assay including all exons from 130 genes involved in cardiovascular Mendelian disorders and analysed simultaneously four samples by multiplexing. Two patients had familial hypertrophic cardiomyopathy (HCM) and two patients suffered from long QT syndrome (LQTS). In patient 1 with HCM, we identified two known pathogenic missense variants in the two most frequently mutated sarcomeric genes MYH7 and MYBPC. In patient 2 with HCM, a known acceptor splice site variant in MYBPC3 was found. In patient 3 with LQTS, two missense variants in the genes SCN5A and KCNQ were identified. Finally, in patient 4 with LQTS a known missense variant was found in MYBPC3, which is usually mutated in patients with cardiomyopathy. Our results showed that multiplex targeted HTS works as an efficient and cost-effective tool for molecular diagnosis of heterogeneous disorders in clinical practice and offers new insights in the pathogenesis of these complex diseases.
