title: Identification of gene mutations in autosomal dominant polycystic kidney disease through targeted resequencing.
abstract: Mutations in two large multi-exon genes, PKD1 and PKD2, cause autosomal dominant polycystic kidney disease (ADPKD). The duplication of PKD1 exons 1-32 as six pseudogenes on chromosome 16, the high level of allelic heterogeneity, and the cost of Sanger sequencing complicate mutation analysis, which can aid diagnostics of ADPKD. We developed and validated a strategy to analyze both the PKD1 and PKD2 genes using next-generation sequencing by pooling long-range PCR amplicons and multiplexing bar-coded libraries. We used this approach to characterize a cohort of 230 patients with ADPKD. This process detected definitely and likely pathogenic variants in 115 (63%) of 183 patients with typical ADPKD. In addition, we identified atypical mutations, a gene conversion, and one missed mutation resulting from allele dropout, and we characterized the pattern of deep intronic variation for both genes. In summary, this strategy involving next-generation sequencing is a model for future genetic characterization of large ADPKD populations.
authors.affiliation: Division of Nephrology and Hypertension, Mayo Clinic, Rochester, MN 55905, USA. rossetti.sandro@mayo.edu
authors.forename: Sandro
authors.initials: S
authors.lastname: Rossetti
authors.forename: Katharina
authors.initials: K
authors.lastname: Hopp
authors.forename: Robert A
authors.initials: RA
authors.lastname: Sikkink
authors.forename: Jamie L
authors.initials: JL
authors.lastname: Sundsbak
authors.forename: Yean Kit
authors.initials: YK
authors.lastname: Lee
authors.forename: Vickie
authors.initials: V
authors.lastname: Kubly
authors.forename: Bruce W
authors.initials: BW
authors.lastname: Eckloff
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Ward
authors.forename: Christopher G
authors.initials: CG
authors.lastname: Winearls
authors.forename: Vicente E
authors.initials: VE
authors.lastname: Torres
authors.forename: Peter C
authors.initials: PC
authors.lastname: Harris
mesh_terms: D001330:Electronic Data Processing; D006801:Humans; D009154:Mutation; D016891:Polycystic Kidney, Autosomal Dominant / Q000235:genetics*; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA / Q000379:methods*; D050396:TRPP Cation Channels / Q000235:genetics*
chemical_list: D050396:TRPP Cation Channels