Non-invasive prenatal diagnosis of achondroplasia and thanatophoric dysplasia: next-generation sequencing allows for a safer, more accurate, and comprehensive approach.
OBJECTIVE
Accurate prenatal diagnosis of genetic conditions can be challenging and usually requires invasive testing. Here, we demonstrate the potential of next-generation sequencing (NGS) for the analysis of cell-free DNA in maternal blood to transform prenatal diagnosis of monogenic disorders.


METHODS
Analysis of cell-free DNA using a PCR and restriction enzyme digest (PCR-RED) was compared with a novel NGS assay in pregnancies at risk of achondroplasia and thanatophoric dysplasia.


RESULTS
PCR-RED was performed in 72 cases and was correct in 88.6%, inconclusive in 7% with one false negative. NGS was performed in 47 cases and was accurate in 96.2% with no inconclusives. Both approaches were used in 27 cases, with NGS giving the correct result in the two cases inconclusive with PCR-RED.


CONCLUSIONS
NGS provides an accurate, flexible approach to non-invasive prenatal diagnosis of de novo and paternally inherited mutations. It is more sensitive than PCR-RED and is ideal when screening a gene with multiple potential pathogenic mutations. These findings highlight the value of NGS in the development of non-invasive prenatal diagnosis for other monogenic disorders.
UCL Institute of Child Health, Genetics and Genomic Medicine, London, UK.|University College London Hospitals NHS Foundation Trust, London, UK.
Lyn S
LS
Chitty
N-E Thames Regional Genetics Service, Great Ormond Street Hospital for Children NHS Foundation Trust, London, UK.
Sarah
S
Mason
N-E Thames Regional Genetics Service, Great Ormond Street Hospital for Children NHS Foundation Trust, London, UK.
Angela N
AN
Barrett
N-E Thames Regional Genetics Service, Great Ormond Street Hospital for Children NHS Foundation Trust, London, UK.
Fiona
http://orcid.org/0000-0003-3626-764X
F
McKay
N-E Thames Regional Genetics Service, Great Ormond Street Hospital for Children NHS Foundation Trust, London, UK.
Nicholas
N
Lench
University College London Hospitals NHS Foundation Trust, London, UK.
Rebecca
R
Daley
N-E Thames Regional Genetics Service, Great Ormond Street Hospital for Children NHS Foundation Trust, London, UK.
Lucy A
LA
Jenkins
D000130:Achondroplasia / Q000175:diagnosis* / Q000235:genetics; D015415:Biomarkers / Q000097:blood; D004247:DNA / Q000097:blood*; D005188:False Negative Reactions; D005260:Female; D005819:Genetic Markers; D005820:Genetic Testing / Q000379:methods*; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D008297:Male; D062145:Maternal Serum Screening Tests; D009154:Mutation; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D051498:Receptor, Fibroblast Growth Factor, Type 3 / Q000235:genetics; D012189:Retrospective Studies; D017422:Sequence Analysis, DNA / Q000379:methods*; D013796:Thanatophoric Dysplasia / Q000175:diagnosis* / Q000235:genetics
D015415:Biomarkers; D005819:Genetic Markers; D004247:DNA; C496365:FGFR3 protein, human; D051498:Receptor, Fibroblast Growth Factor, Type 3