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Hum Mutat. 2017 Jul;38(7):870-879. doi: 10.1002/humu.23223. Epub 2017 May 29.

Detecting PKD1 variants in polycystic kidney disease patients by single-molecule long-read sequencing.

Author information

GenomeScan B.V, Leiden, The Netherlands.
Institut National de la Santé et de la Recherche Médicale (INSERM), Institut of Cardiovascular and Metabolic Disease, Toulouse, France.
Université Toulouse III Paul-Sabatier, Toulouse, France.
Leiden Genome Technology Center (LGTC), Department of Human Genetics, Leiden University Medical Center (LUMC), Leiden, The Netherlands.
Department of Human Genetics, Leiden University Medical Center (LUMC), Leiden, The Netherlands.
Department of Nephrology, University Hospital Groningen, University Medical Center Groningen, Groningen, The Netherlands.
Department of Clinical Genetics, Leiden University Medical Center (LUMC), Leiden, The Netherlands.


A genetic diagnosis of autosomal-dominant polycystic kidney disease (ADPKD) is challenging due to allelic heterogeneity, high GC content, and homology of the PKD1 gene with six pseudogenes. Short-read next-generation sequencing approaches, such as whole-genome sequencing and whole-exome sequencing, often fail at reliably characterizing complex regions such as PKD1. However, long-read single-molecule sequencing has been shown to be an alternative strategy that could overcome PKD1 complexities and discriminate between homologous regions of PKD1 and its pseudogenes. In this study, we present the increased power of resolution for complex regions using long-read sequencing to characterize a cohort of 19 patients with ADPKD. Our approach provided high sensitivity in identifying PKD1 pathogenic variants, diagnosing 94.7% of the patients. We show that reliable screening of ADPKD patients in a single test without interference of PKD1 homologous sequences, commonly introduced by residual amplification of PKD1 pseudogenes, by direct long-read sequencing is now possible. This strategy can be implemented in diagnostics and is highly suitable to sequence and resolve complex genomic regions that are of clinical relevance.


ADPKD; DNA diagnostics; PKD1; PacBio; complex genomic regions; long-read sequencing; single-molecule real-time sequencing; variant detection

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