title: High-density oligonucleotide-based resequencing assay for mutations causing syndromic and non-syndromic forms of thoracic aortic aneurysms and dissections.
abstract: Thoracic aortic aneurysm and dissection is associated with increasing mortality rate that may occur as part of a syndrome or as an isolated familial condition. Several genes have been implicated in causing TAAD, though an appropriate genetic test for their parallel testing is not yet available. Herein, we describe the novel 117-kb "MFSTAAD chip" that may help to understand the genetic basis of TAAD. A custom duplicate resequencing assay was developed to cover eight genes previously described in TAAD; FBN1, TGFBR1&2, COL3A1, MYH11, ACTA2, SLC2A10 and NOTCH1. GSEQ and SeqC software were used for data analysis. The analytical sensitivity of the assay was validated by the recognition of 182 known mutations (153 point mutations, 21 deletions, 7 insertions and 1 duplication) and a cohort of 28 patients were selected to determine the mutation yield, whereby 18 of them were previously negative for mutations in the genes FBN1 and TGFBR2. The assay had significantly higher sensitivity for point mutations (100%) and the largest deletion of 16 bp was detectable through a decline in the hybridization strength. The overall analytical sensitivity was 85%. Mutation testing of 28 unrelated TAAD patients revealed 4 known and 6 possibly pathogenic mutations with a mutation yield of 32%. The MFSTAAD chip is an alternative tool to next-generation sequencing that allows parallel analysis of several genes on a single platform. Refinements in the probe design and data analysis software will increase the analytical sensitivity of insertions and deletions making this assay even more applicable for clinical testing.
authors.affiliation: Institute for Human Genetics, Medical School of Hannover, Carl-Neuberg-Str. 1, 30625 Hannover, Germany.
authors.forename: Ushanthine
authors.initials: U
authors.lastname: Kathiravel
authors.forename: Britta
authors.initials: B
authors.lastname: Keyser
authors.forename: Sabine
authors.initials: S
authors.lastname: Hoffjan
authors.forename: Judith
authors.initials: J
authors.lastname: Kötting
authors.forename: Melanie
authors.initials: M
authors.lastname: Müller
authors.forename: Sugirthan
authors.initials: S
authors.lastname: Sivalingam
authors.forename: Michael
authors.initials: M
authors.lastname: Bonin
authors.forename: Mine
authors.initials: M
authors.lastname: Arslan-Kirchner
authors.forename: Yskert
authors.initials: Y
authors.lastname: von Kodolitsch
authors.forename: Priska
authors.initials: P
authors.lastname: Binner
authors.forename: Thomas
authors.initials: T
authors.lastname: Scheffold
authors.forename: Manfred
authors.initials: M
authors.lastname: Stuhrmann
authors.forename: Stephan
authors.initials: S
authors.lastname: Waldmüller
mesh_terms: D000784:Aortic Dissection / Q000235:genetics*; D017545:Aortic Aneurysm, Thoracic / Q000235:genetics*; D001483:Base Sequence; D003198:Computer Simulation; D004252:DNA Mutational Analysis / Q000295:instrumentation / Q000379:methods*; D005189:False Positive Reactions; D000071838:Fibrillin-1; D000071837:Fibrillins; D020022:Genetic Predisposition to Disease; D006801:Humans; D008840:Microfilament Proteins / Q000235:genetics; D008969:Molecular Sequence Data; D009154:Mutation; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D009841:Oligonucleotides; D012680:Sensitivity and Specificity; D054392:Adipokines
chemical_list: D000071838:Fibrillin-1; D000071837:Fibrillins; D008840:Microfilament Proteins; D009841:Oligonucleotides; C000605722:FBN1 protein, human; D054392:Adipokines