Sensitive Monogenic Noninvasive Prenatal Diagnosis by Targeted Haplotyping

Am J Hum Genet. 2017 Sep 7;101(3):326-339. doi: 10.1016/j.ajhg.2017.07.012. Epub 2017 Aug 24.


During pregnancy, cell-free DNA (cfDNA) in maternal blood encompasses a small percentage of cell-free fetal DNA (cffDNA), an easily accessible source for determination of fetal disease status in risk families through non-invasive procedures. In case of monogenic heritable disease, background maternal cfDNA prohibits direct observation of the maternally inherited allele. Non-invasive prenatal diagnostics (NIPD) of monogenic diseases therefore relies on parental haplotyping and statistical assessment of inherited alleles from cffDNA, techniques currently unavailable for routine clinical practice. Here, we present monogenic NIPD (MG-NIPD), which requires a blood sample from both parents, for targeted locus amplification (TLA)-based phasing of heterozygous variants selectively at a gene of interest. Capture probes-based targeted sequencing of cfDNA from the pregnant mother and a tailored statistical analysis enables predicting fetal gene inheritance. MG-NIPD was validated for 18 pregnancies, focusing on CFTR, CYP21A2, and HBB. In all cases we could predict the inherited alleles with >98% confidence, even at relatively early stages (8 weeks) of pregnancy. This prediction and the accuracy of parental haplotyping was confirmed by sequencing of fetal material obtained by parallel invasive procedures. MG-NIPD is a robust method that requires standard instrumentation and can be implemented in any clinic to provide families carrying a severe monogenic disease with a prenatal diagnostic test based on a simple blood draw.

Keywords: CAH; NIPD; TLA; beta thalassemia; cell-free DNA; cfDNA; congenital adrenal hyperplasia; cystic fibrosis; monogenic diseases; non-invasive prenatal diagnosis; targeted haplotyping; targeted locus amplification.

MeSH terms

  • Adrenal Hyperplasia, Congenital / blood
  • Adrenal Hyperplasia, Congenital / diagnosis*
  • Adrenal Hyperplasia, Congenital / genetics
  • Biomarkers / blood*
  • Cells, Cultured
  • Cystic Fibrosis / blood
  • Cystic Fibrosis / diagnosis*
  • Cystic Fibrosis / genetics
  • Cystic Fibrosis Transmembrane Conductance Regulator / blood
  • Cystic Fibrosis Transmembrane Conductance Regulator / genetics*
  • DNA / blood
  • DNA / genetics
  • Female
  • Haplotypes
  • Humans
  • Polymorphism, Single Nucleotide*
  • Pregnancy
  • Prenatal Diagnosis / methods*
  • Steroid 21-Hydroxylase / blood
  • Steroid 21-Hydroxylase / genetics*


  • Biomarkers
  • CFTR protein, human
  • Cystic Fibrosis Transmembrane Conductance Regulator
  • DNA
  • CYP21A2 protein, human
  • Steroid 21-Hydroxylase