Allelic dropout in PAH affecting the results of genetic diagnosis in phenylketonuria

J Pediatr Endocrinol Metab. 2022 Jan 14;35(3):387-391. doi: 10.1515/jpem-2021-0336. Print 2022 Mar 28.

Abstract

Objectives: Phenylketonuria (PKU) is an inherited autosomal recessive disorder of phenylalanine metabolism. It is mainly caused by a deficiency in phenylalanine hydroxylase (PAH) and frequently diagnosed with Sanger sequencing. To some extent, allelic dropout can explain the inconsistency in genotype and phenotype.

Methods: Three families were evaluated through DNA sequence analysis, multiplex ligation-dependent probe amplification (MLPA) and prenatal diagnosis technologies. The possibility of inconsistency in phenotype and genotype with c.331C>T variant was analysed.

Results: Through pedigree analysis, three mothers carried a homozygous c.331C>T variant, which was a false-positive result. New primers were used, and this error was caused by allelic dropout. In this case, c.158G>A was likely a benign variant.

Conclusions: Sequence variants in primer-binding regions could cause allelic dropout, creating unpredictable errors in genotyping. Our results emphasised the need for careful measures to treat genotype-phenotype inconsistencies.

Keywords: allelic dropout; phenylketonuria (PKU); variant.

MeSH terms

  • Alleles
  • Female
  • Genotype
  • Humans
  • Mutation
  • Phenotype
  • Phenylalanine Hydroxylase* / genetics
  • Phenylalanine Hydroxylase* / metabolism
  • Phenylketonurias* / diagnosis
  • Phenylketonurias* / genetics
  • Pregnancy

Substances

  • Phenylalanine Hydroxylase