Noninvasive prenatal test of single-gene disorders by linked-read direct haplotyping: application in various diseases


Direct haplotyping enables noninvasive prenatal testing (NIPT) without analyzing proband, which is a promising strategy for pregnancies at risk of an inherited single-gene disorder. Here, we aimed to expand the scope of single-gene disorders that NIPT using linked-read direct haplotyping would be applicable to. Three families at risk of myotonic dystrophy type 1, lipoid congenital adrenal hyperplasia, and Fukuyama congenital muscular dystrophy were recruited. All cases exhibited distinct characteristics that are often encountered as hurdles (i.e., repeat expansion, identical variants in both parents, and novel variants with retrotransposon insertion) in the universal clinical application of NIPT. Direct haplotyping of parental genomes was performed by linked-read sequencing, combined with allele-specific PCR, if necessary. Target DMPK, STAR, and FKTN genes in the maternal plasma DNA were sequenced. Posterior risk calculations and an Anderson–Darling test were performed to deduce the maternal and paternal inheritance, respectively. In all cases, we could predict the inheritance of maternal mutant allele with > 99.9% confidence, while paternal mutant alleles were not predicted to be inherited. Our study indicates that direct haplotyping and posterior risk calculation can be applied with subtle modifications to NIPT for the detection of an expanded range of diseases.

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Fig. 1: Flow chart of the overall strategy for noninvasive prenatal diagnosis of single-gene disorders in this study.
Fig. 2: Pedigree of three families with DM1 (family A), Lipoid CAH (family B), and FCMD (family C).
Fig. 3: Distribution of the fraction of Hap I alleles according to the inheritance of either maternal Hap I or Hap II in three families with DM1 (A), Lipoid CAH (B), and FCMD (C).


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This study was supported by grant 03-2016-0050 from the Seoul National University Hospital (SNUH) Research Fund.

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Correspondence to Moon-Woo Seong.

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Lee, JS., Lee, K.B., Song, H. et al. Noninvasive prenatal test of single-gene disorders by linked-read direct haplotyping: application in various diseases. Eur J Hum Genet (2020).

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