Noninvasive prenatal test for FGFR3-related skeletal dysplasia based on next-generation sequencing and plasma cell-free DNA: Test performance analysis and feasibility exploration.

Abstract

OBJECTIVE:

To explore the feasibility and accuracy of a noninvasive prenatal test for fibroblast growth factor receptor 3 (FGFR3)-related skeletal dysplasia based on next-generation sequencing (NGS) of plasma cell-free DNA.

METHOD:

Fragmented genome DNA (gDNA) of fetuses with achondroplasia (ACH) and thanatophoric dysplasia type I (TD I) was mixed with postdelivery maternal plasma cell-free DNA to generate spiked samples of different modeled fetal fractions. Multiplex polymerase chain reaction was used to amplify the 19 FGFR3 loci, and the amplification products were then sequenced by NGS to detect the fetal mutant alleles. Then, maternal plasma samples of pregnant women carrying ACH (n = 4) and TD I fetuses (n = 2), as well as healthy controls (n = 15), were tested by NGS, and the test performance was evaluated.

RESULTS:

Fetal FGFR3 mutations were detected in all artificial mixtures with fetal gDNA concentrations above 3%. In clinical validation, our method identified all fetal FGFR3 mutant alleles from maternal plasma, with no false positive results. The sensitivity and specificity of our method were 100% (95% CI, 54.1%-100%) and 100% (78.2%-100%), respectively.

CONCLUSION:

Our method had a favorable performance for noninvasively detecting fetal FGFR3 mutations in maternal plasma, highlighting its promising value in developing a noninvasive prenatal test for de novo and paternally inherited disorders.

• Authors: Yuan Ren, Jia Zhao, Ruibing Li, Yifan Xie, Shufang Jiang, Honghui Zhou, Hongtai Liu, Yanqin You, Fang Chen, Wei Wang, Ya Gao, Yuanguang Meng, Yanping Lu
• Journal: Prenatal diagnosis (Prenat Diagn) Vol. 38 Issue 11 Pg. 821-828 (10 2018) ISSN: 1097-0223 [Electronic] England
• PMID: 30048571 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't, Validation Study)
• Copyright: © 2018 John Wiley & Sons, Ltd.

Chemical References

• Cell-Free Nucleic Acids
• FGFR3 protein, human
• Receptor, Fibroblast Growth Factor, Type 3

Topics

• Case-Control Studies
• Cell-Free Nucleic Acids (analysis)
• Feasibility Studies
• Female
• Humans
• Maternal Serum Screening Tests
• Multiplex Polymerase Chain Reaction
• Pregnancy
• Receptor, Fibroblast Growth Factor, Type 3 (genetics)
• Thanatophoric Dysplasia (diagnosis, genetics)