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Comparative Study
. 2015 Jul;35(7):656-62.
doi: 10.1002/pd.4583. Epub 2015 May 26.

Non-invasive prenatal diagnosis of achondroplasia and thanatophoric dysplasia: next-generation sequencing allows for a safer, more accurate, and comprehensive approach

Lyn S Chitty  1   2 , Sarah Mason  3 , Angela N Barrett  3 , Fiona McKay  3 , Nicholas Lench  3 , Rebecca Daley  2 , Lucy A Jenkins  3
Affiliations
Comparative Study

Non-invasive prenatal diagnosis of achondroplasia and thanatophoric dysplasia: next-generation sequencing allows for a safer, more accurate, and comprehensive approach

Lyn S Chitty et al. Prenat Diagn. 2015 Jul.

Abstract

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.

Conclusion: 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.

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Figures

Figure 1
Figure 1
PCR-restriction enzyme digest results for (a) achondroplasia [fibroblast growth factor receptor 3 (FGFR3): c.1138G>A p.(Gly380Arg)/restriction enzyme BsrG1] and (b) thanatophoric dysplasia [FGFR3: c.742C>T p.(Arg248Cys)/restriction enzyme DraIII]. Upper arrows indicate wild-type (normal) allele that is strongly present in all samples. Bottom arrows indicate the mutant alleles that are faint in affected cell-free DNA (cfDNA+) and stronger mutation positive control genomic DNA (gDNA+ve). There is no mutant band present in the unaffected maternal genomic DNA (Mat gDNA). The figure shows uncropped images visualized on Shimadzu Multi-NA microchip electrophoresis system. NTC, blank, no template control
See this image and copyright information in PMC

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