title: Target enrichment and high-throughput sequencing of 80 ribosomal protein genes to identify mutations associated with Diamond-Blackfan anaemia.
abstract: Diamond-Blackfan anaemia (DBA) is caused by inactivating mutations in ribosomal protein (RP) genes, with mutations in 13 of the 80 RP genes accounting for 50-60% of cases. The remaining 40-50% cases may harbour mutations in one of the remaining RP genes, but the very low frequencies render conventional genetic screening as challenging. We, therefore, applied custom enrichment technology combined with high-throughput sequencing to screen all 80 RP genes. Using this approach, we identified and validated inactivating mutations in 15/17 (88%) DBA patients. Target enrichment combined with high-throughput sequencing is a robust and improved methodology for the genetic diagnosis of DBA.
authors.affiliation: Imperial Molecular Pathology Laboratory, Imperial College Healthcare NHS Trust and Academic Health Sciences Centre, Hammersmith Hospital, London, UK. g.gerrard@imperial.ac.uk
authors.forename: Gareth
authors.initials: G
authors.lastname: Gerrard
authors.forename: Mikel
authors.initials: M
authors.lastname: Valgañón
authors.forename: Hui En
authors.initials: HE
authors.lastname: Foong
authors.forename: Dalia
authors.initials: D
authors.lastname: Kasperaviciute
authors.forename: Deena
authors.initials: D
authors.lastname: Iskander
authors.forename: Laurence
authors.initials: L
authors.lastname: Game
authors.forename: Michael
authors.initials: M
authors.lastname: Müller
authors.forename: Timothy J
authors.initials: TJ
authors.lastname: Aitman
authors.forename: Irene
authors.initials: I
authors.lastname: Roberts
authors.forename: Josu
authors.initials: J
authors.lastname: de la Fuente
authors.forename: Letizia
authors.initials: L
authors.lastname: Foroni
authors.forename: Anastasios
authors.initials: A
authors.lastname: Karadimitris
mesh_terms: D000293:Adolescent; D000328:Adult; D029503:Anemia, Diamond-Blackfan / Q000235:genetics*; D002648:Child; D002675:Child, Preschool; D018389:Codon, Nonsense; D004252:DNA Mutational Analysis; D004275:DNA, Ribosomal / Q000097:blood / Q000235:genetics*; D005260:Female; D016368:Frameshift Mutation; D015723:Gene Library; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D008297:Male; D012269:Ribosomal Proteins / Q000235:genetics*; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017384:Sequence Deletion
chemical_list: D018389:Codon, Nonsense; D004275:DNA, Ribosomal; D012269:Ribosomal Proteins
keywords: Diamond-Blackfan anaemia; molecular diagnostics; next generation sequencing; target enrichment