title: Neurofibromatosis type 1 gene mutation analysis using sequence capture and high-throughput sequencing.
abstract: Neurofibromatosis type 1 syndrome (NF1) is caused by mutations in the NF1 gene. Availability of new sequencing technology prompted us to search for an alternative method for NF1 mutation analysis. Genomic DNA was isolated from saliva avoiding invasive sampling. The NF1 exons with an additional 50bp of flanking intronic sequences were captured and enriched using the SeqCap EZ Choice Library protocol. The captured DNA was sequenced with the Roche/454 GS Junior system. The mean coverages of the targeted regions were 41x and 74x in 2 separate sets of samples. An NF1 mutation was discovered in 10 out of 16 separate patient samples. Our study provides proof of principle that the sequence capture methodology combined with high-throughput sequencing is applicable to NF1 mutation analysis. Deep intronic mutations may however remain undetectable, and change at the DNA level may not predict the outcome at the mRNA or protein levels.
authors.affiliation: Department of Cell Biology and Anatomy, University of Turku, Turku, Finland.
authors.forename: Elina
authors.initials: E
authors.lastname: Uusitalo
authors.forename: Anna
authors.initials: A
authors.lastname: Hammais
authors.forename: Elina
authors.initials: E
authors.lastname: Palonen
authors.forename: Annika
authors.initials: A
authors.lastname: Brandt
authors.forename: Ville-Veikko
authors.initials: VV
authors.lastname: Mäkelä
authors.forename: Roope
authors.initials: R
authors.lastname: Kallionpää
authors.forename: Eeva-Mari
authors.initials: EM
authors.lastname: Jouhilahti
authors.forename: Minna
authors.initials: M
authors.lastname: Pöyhönen
authors.forename: Juhani
authors.initials: J
authors.lastname: Soini
authors.forename: Juha
authors.initials: J
authors.lastname: Peltonen
authors.forename: Sirkku
authors.initials: S
authors.lastname: Peltonen
mesh_terms: D004252:DNA Mutational Analysis / Q000379:methods*; D005091:Exons; D016514:Genes, Neurofibromatosis 1; D020022:Genetic Predisposition to Disease; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D006801:Humans; D007438:Introns; D009154:Mutation; D009456:Neurofibromatosis 1 / Q000175:diagnosis / Q000235:genetics*; D011237:Predictive Value of Tests; D012463:Saliva / Q000737:chemistry