Haplotype-based approach for noninvasive prenatal diagnosis of congenital adrenal hyperplasia by maternal plasma DNA sequencing

Gene. 2014 Jul 10;544(2):252-8. doi: 10.1016/j.gene.2014.04.055. Epub 2014 Apr 24.

Abstract

Prenatal diagnosis of congenital adrenal hyperplasia (CAH) is of clinical significance because in utero treatment is available to prevent virilization of an affected female fetus. However, traditional prenatal diagnosis of CAH relies on genetic testing of fetal genomic DNA obtained using amniocentesis or chorionic villus sampling, which is associated with an increased risk of miscarriage. The aim of this study was to demonstrate the feasibility of a new haplotype-based approach for the noninvasive prenatal testing of CAH due to 21-hydroxylase deficiency. Parental haplotypes were constructed using target-region sequencing data of the parents and the proband. With the assistance of the parental haplotypes, we recovered fetal haplotypes using a hidden Markov model (HMM) through maternal plasma DNA sequencing. In the genomic region around the CYP21A2 gene, the fetus inherited the paternal haplotype '0' alleles linked to the mutant CYP21A2 gene, but the maternal haplotype '1' alleles linked to the wild-type gene. The fetus was predicted to be an unaffected carrier of CAH, which was confirmed by genetic analysis of fetal genomic DNA from amniotic fluid cells. This method was further validated by comparing the inferred SNP genotypes with the direct sequencing data of fetal genomic DNA. The result showed an accuracy of 96.41% for the inferred maternal alleles and an accuracy of 97.81% for the inferred paternal alleles. The haplotype-based approach is feasible for noninvasive prenatal testing of CAH.

Keywords: Congenital adrenal hyperplasia; Fetal DNA in maternal plasma; Haplotype; Massively parallel sequencing; Noninvasive prenatal testing.

Publication types

  • Case Reports
  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adrenal Hyperplasia, Congenital / diagnosis*
  • Adrenal Hyperplasia, Congenital / genetics*
  • Amniotic Fluid / cytology
  • DNA / blood
  • DNA / genetics
  • Female
  • Genetic Testing / methods*
  • Genotype
  • Haplotypes
  • Humans
  • Male
  • Polymorphism, Genetic
  • Pregnancy
  • Prenatal Diagnosis / methods*
  • Sequence Analysis, DNA / methods
  • Steroid 21-Hydroxylase / blood
  • Steroid 21-Hydroxylase / genetics

Substances

  • DNA
  • CYP21A2 protein, human
  • Steroid 21-Hydroxylase

Supplementary concepts

  • Congenital adrenal hyperplasia due to 21 hydroxylase deficiency