Molecular characterization, by digital PCR analysis of four HMBS gene mutations affecting the ubiquitous isoform of Porphobilinogen Deaminase (PBGD) in patients with Acute Intermittent Porphyria (AIP)

Mol Genet Metab. 2018 Nov;125(3):295-301. doi: 10.1016/j.ymgme.2018.09.002. Epub 2018 Sep 5.

Abstract

Genetic variants in promoters and alternative-splicing lesions require to be experimentally tested in order to validate them as causatives of a disease. The digital PCR (dPCR) approach, which is an alternative to the classical qPCR, is an innovative and a more sensitive method for the detection and quantification of nucleic acids. In the present study, we identified four HMBS gene mutations affecting the ubiquitous isoform of porphobilinogen deaminase (PBGD) and established a dPCR protocol which would be able to detect the different transcripts of this gene. With the application of this method, we were able to characterize the functional roles of these four genetic variants, demonstrating that all these mutations were causatives of the non-erythroid variant of the acute intermittent porphyria (AIP) disease.

Keywords: Digital PCR; Gene expression; HMBS; Porphyria; Promoter variants; Splicing isoform.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Child
  • Female
  • Humans
  • Hydroxymethylbilane Synthase / genetics*
  • Male
  • Middle Aged
  • Mutation
  • Polymerase Chain Reaction
  • Porphyria, Acute Intermittent / genetics*
  • Porphyria, Acute Intermittent / pathology
  • Promoter Regions, Genetic / genetics
  • Protein Isoforms / genetics*
  • RNA Splice Sites / genetics*
  • Young Adult

Substances

  • Protein Isoforms
  • RNA Splice Sites
  • Hydroxymethylbilane Synthase