Mutation analyses by next-generation sequencing and multiplex ligation-dependent probe amplification in Japanese autosomal dominant polycystic kidney disease patients

Clin Exp Nephrol. 2019 Aug;23(8):1022-1030. doi: 10.1007/s10157-019-01736-3. Epub 2019 Apr 15.

Abstract

Background: Autosomal dominant polycystic kidney disease (ADPKD), one of the most common hereditary kidney diseases, causes gradual growth of cysts in the kidneys, leading to renal failure. Owing to the advanced technology of next-generation sequencing (NGS), genetic analyses of the causative genes PKD1 and PKD2 have been improved.

Methods: We performed genetic analyses of 111 Japanese ADPKD patients using hybridization-based NGS and long-range (LR)-PCR-based NGS. Additionally, genetic analyses in exon 1 of PKD1 using Sanger sequencing because of an extremely low coverage of NGS and those using multiplex ligation-dependent probe amplification (MLPA) were performed.

Results: The detection rate using NGS for 111 patients was 86.5%. One mutation in exon 1 of PKD1 and five deletions detected by MLPA were identified. When combined, the total detection rate was 91.9%.

Conclusion: Although NGS is useful, we propose the addition of Sanger sequencing for exon 1 of PKD1 and MLPA as indispensable for identifying mutations not detected by NGS.

Keywords: ADPKD; MLPA; Next-generation sequencing; PKD1; PKD2; Polycystic kidney disease.

MeSH terms

  • Asian People / genetics
  • DNA Mutational Analysis / methods*
  • Exons
  • Genetic Predisposition to Disease
  • High-Throughput Nucleotide Sequencing*
  • Humans
  • Japan
  • Multiplex Polymerase Chain Reaction*
  • Mutation*
  • Polycystic Kidney, Autosomal Dominant / diagnosis
  • Polycystic Kidney, Autosomal Dominant / ethnology
  • Polycystic Kidney, Autosomal Dominant / genetics*
  • Predictive Value of Tests
  • Risk Factors
  • TRPP Cation Channels / genetics*

Substances

  • TRPP Cation Channels
  • polycystic kidney disease 1 protein