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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
Next-generation sequencing using microfluidic PCR enrichment for molecular autopsy
BMC Cardiovascular Disorders, Volume 19, No. 1, Article 174, Year 2019
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Description
Background: We aimed to determine the mutation yield and clinical applicability of "molecular autopsy" following sudden arrhythmic death syndrome (SADS) by validating and utilizing low-cost high-throughput technologies: Fluidigm Access Array PCR-enrichment with Illumina HiSeq 2000 next generation sequencing (NGS). Methods: We validated and optimized the NGS platform with a subset of 46 patients by comparison with Sanger sequencing of coding exons of major arrhythmia risk-genes (KCNQ1, KCNH2, SCN5A, KCNE1, KCNE2, RYR2). A combined large multi-ethnic international SADS cohort was sequenced utilizing the NGS platform to determine overall molecular yield; rare variants identified by NGS were subsequently reconfirmed by Sanger sequencing. Results: The NGS platform demonstrated 100% sensitivity for pathogenic variants as well as 87.20% sensitivity and 99.99% specificity for all substitutions (optimization subset, n = 46). The positive predictive value (PPV) for NGS for rare substitutions was 16.0% (27 confirmed rare variants of 169 positive NGS calls in 151 additional cases). The overall molecular yield in 197 multi-ethnic SADS cases (mean age 22.6 ± 14.4 years, 68% male) was 5.1% (95% confidence interval 2.0-8.1%), representing 10 cases carrying pathogenic or likely pathogenic risk-mutations. Conclusions: Molecular autopsy with Fluidigm Access Array and Illumina HiSeq NGS utilizing a selected panel of LQTS/BrS and CPVT risk-genes offers moderate diagnostic yield, albeit requiring confirmatory Sanger-sequencing of mutational variants. © 2019 The Author(s).
Authors & Co-Authors
Raju, Hariharan
United Kingdom, London
St George’s, University of London
Australia, Macquarie Park
The Faculty of Medicine, Health and Human Sciences
Ware, James S.
United Kingdom, London
Royal Brompton Hospital
Skinner, Jonathan Robert
New Zealand, Auckland
Starship Children’s Hospital
Hedley, P. L.
Denmark, Copenhagen
Statens Serum Institut
Arno, Gavin
United Kingdom, London
St George’s, University of London
Love, Donald R.
New Zealand, Auckland
Auckland City Hospital
van der Werf, Christian
Netherlands, Amsterdam
Amsterdam Cardiovascular Sciences
Tfelt-Hansen, Jacob
Denmark, Copenhagen
Copenhagen University Hospital
Denmark, Copenhagen
Københavns Universitet
Belgium, Brussels
European Union
Winkel, Bo Gregers
Denmark, Copenhagen
Copenhagen University Hospital
Belgium, Brussels
European Union
Cohen, M. C.
United Kingdom, Sheffield
Sheffield Children's Hospital
Li, Xinzhong
United Kingdom, London
Royal Brompton Hospital
United Kingdom, Middlesbrough
Teesside University
John, Shibu
United Kingdom, London
Royal Brompton Hospital
Sharma, Sanjay Sanjay
United Kingdom, London
St George’s, University of London
Jeffery, Stephen C.C.
United Kingdom, London
St George’s, University of London
Wilde, Arthur A.M.
Netherlands, Amsterdam
Amsterdam Cardiovascular Sciences
Belgium, Brussels
European Union
Christiansen, Michael
Denmark, Copenhagen
Statens Serum Institut
Denmark, Copenhagen
Københavns Universitet
Sheppard, M. N.
United Kingdom, London
St George’s, University of London
Belgium, Brussels
European Union
Behr, Elijah R.
United Kingdom, London
St George’s, University of London
Belgium, Brussels
European Union
Statistics
Citations: 6
Authors: 18
Affiliations: 12
Identifiers
Doi:
10.1186/s12872-019-1154-8
ISSN:
14712261
Research Areas
Cancer
Health System And Policy
Study Design
Cohort Study
Participants Gender
Male