dc.contributor.author |
Raju, Hariharan |
en |
dc.contributor.author |
Ware, James S |
en |
dc.contributor.author |
Skinner, Jonathan |
en |
dc.contributor.author |
Hedley, Paula L |
en |
dc.contributor.author |
Arno, Gavin |
en |
dc.contributor.author |
Love, Donald |
en |
dc.contributor.author |
van der Werf, Christian |
en |
dc.contributor.author |
Tfelt-Hansen, Jacob |
en |
dc.contributor.author |
Winkel, Bo Gregers |
en |
dc.contributor.author |
Cohen, Marta C |
en |
dc.contributor.author |
Li, Xinzhong |
en |
dc.contributor.author |
John, Shibu |
en |
dc.contributor.author |
Sharma, Sanjay |
en |
dc.contributor.author |
Jeffery, Steve |
en |
dc.contributor.author |
Wilde, Arthur AM |
en |
dc.contributor.author |
Christiansen, Michael |
en |
dc.contributor.author |
Sheppard, Mary N |
en |
dc.contributor.author |
Behr, Elijah R |
en |
dc.date.accessioned |
2020-02-07T00:55:38Z |
en |
dc.date.issued |
2019-07-23 |
en |
dc.identifier.citation |
BMC cardiovascular disorders 19(1):174 23 Jul 2019 |
en |
dc.identifier.issn |
1471-2261 |
en |
dc.identifier.uri |
http://hdl.handle.net/2292/49789 |
en |
dc.description.abstract |
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. |
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dc.format.medium |
Electronic |
en |
dc.language |
eng |
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dc.relation.ispartofseries |
BMC cardiovascular disorders |
en |
dc.rights |
Items in ResearchSpace are protected by copyright, with all rights reserved, unless otherwise indicated. Previously published items are made available in accordance with the copyright policy of the publisher. |
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dc.rights.uri |
https://researchspace.auckland.ac.nz/docs/uoa-docs/rights.htm |
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dc.rights.uri |
https://creativecommons.org/licenses/by/4.0/ |
en |
dc.subject |
Humans |
en |
dc.subject |
Death, Sudden, Cardiac |
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dc.subject |
Genetic Predisposition to Disease |
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dc.subject |
Autopsy |
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dc.subject |
Microfluidic Analytical Techniques |
en |
dc.subject |
Cause of Death |
en |
dc.subject |
Risk Factors |
en |
dc.subject |
Reproducibility of Results |
en |
dc.subject |
Predictive Value of Tests |
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dc.subject |
Polymerase Chain Reaction |
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dc.subject |
Pedigree |
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dc.subject |
DNA Mutational Analysis |
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dc.subject |
Heredity |
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dc.subject |
Mutation |
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dc.subject |
Adolescent |
en |
dc.subject |
Adult |
en |
dc.subject |
Child |
en |
dc.subject |
Child, Preschool |
en |
dc.subject |
Infant |
en |
dc.subject |
Australia |
en |
dc.subject |
Europe |
en |
dc.subject |
New Zealand |
en |
dc.subject |
Female |
en |
dc.subject |
Male |
en |
dc.subject |
Arrhythmias, Cardiac |
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dc.subject |
Young Adult |
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dc.subject |
Pathology, Molecular |
en |
dc.subject |
High-Throughput Nucleotide Sequencing |
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dc.title |
Next-generation sequencing using microfluidic PCR enrichment for molecular autopsy. |
en |
dc.type |
Journal Article |
en |
dc.identifier.doi |
10.1186/s12872-019-1154-8 |
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pubs.issue |
1 |
en |
pubs.begin-page |
174 |
en |
pubs.volume |
19 |
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dc.rights.holder |
Copyright: The authors |
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pubs.publication-status |
Published |
en |
dc.rights.accessrights |
http://purl.org/eprint/accessRights/OpenAccess |
en |
pubs.subtype |
Research Support, Non-U.S. Gov't |
en |
pubs.subtype |
research-article |
en |
pubs.subtype |
Validation Study |
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pubs.subtype |
Multicenter Study |
en |
pubs.subtype |
Journal Article |
en |
pubs.elements-id |
779007 |
en |
dc.identifier.eissn |
1471-2261 |
en |
pubs.record-created-at-source-date |
2019-07-25 |
en |
pubs.dimensions-id |
31337358 |
en |