High throughput sequencing identifies p.Y1435X as a novel truncating PKD1 mutation

Abstract

INTRODUCTION: Autosomal dominant polycystic kidney disease (ADPKD), the commonest heritable renal disorder, is caused by mutations in PKD1 or PKD2. PKD1 mutations account for 85% of cases and cause more severe disease with earlier onset of end-stage renal disease (ESRD). Maltese ADPKD patients reach ESRD at an early age, generally between 40 and 50 years.

MATERIALS AND METHODS: Blood and saliva samples were obtained from a family with ADPKD. High throughput sequencing (HTS) was used to sequence the entire PKD1 and PKD2 coding regions extending up to 50 bp into the introns in each direction. SureSelectXT Target Enrichment capture of a 2.6 Mb region, including PKD1 and PKD2, was followed by sequencing on Illumina HiSeq4000. HTS data was mapped to GRCh37 as paired-end libraries using NextGENe software. A BED file was used to ensure mapping to PKD1 and PKD2 and excluding pseudogenes. To remove potential pseudogene contamination of data, the mutation list was filtered against an in-house database of 90 control HTS datasets and by pairwise blast of PKD1 and relevant pseudogenes.

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