TY - JOUR
T1 - Sensitive Monogenic Noninvasive Prenatal Diagnosis by Targeted Haplotyping
AU - Vermeulen, Carlo
AU - Geeven, Geert
AU - de Wit, Elzo
AU - Verstegen, Marjon J A M
AU - Jansen, Rumo P M
AU - van Kranenburg, Melissa
AU - de Bruijn, Ewart
AU - Pulit, Sara L
AU - Kruisselbrink, Evelien
AU - Shahsavari, Zahra
AU - Omrani, Davood
AU - Zeinali, Fatemeh
AU - Najmabadi, Hossein
AU - Katsila, Theodora
AU - Vrettou, Christina
AU - Patrinos, George P
AU - Traeger-Synodinos, Joanne
AU - Splinter, Erik
AU - Beekman, Jeffrey M
AU - Kheradmand Kia, Sima
AU - Te Meerman, Gerard J
AU - Ploos van Amstel, Hans Kristian
AU - de Laat, Wouter
N1 - Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.
PY - 2017/9/7
Y1 - 2017/9/7
N2 - 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.
AB - 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.
KW - Adrenal Hyperplasia, Congenital
KW - Biomarkers
KW - Cells, Cultured
KW - Cystic Fibrosis
KW - Cystic Fibrosis Transmembrane Conductance Regulator
KW - DNA
KW - Female
KW - Haplotypes
KW - Humans
KW - Polymorphism, Single Nucleotide
KW - Pregnancy
KW - Prenatal Diagnosis
KW - Steroid 21-Hydroxylase
KW - Journal Article
U2 - 10.1016/j.ajhg.2017.07.012
DO - 10.1016/j.ajhg.2017.07.012
M3 - Article
C2 - 28844486
SN - 0002-9297
VL - 101
SP - 326
EP - 339
JO - American Journal of Human Genetics
JF - American Journal of Human Genetics
IS - 3
ER -