Combined PGD and PGS by NGS on the same biopsy using a single index

Jasper, Melinda J.; Brockman, M D O Ross; Hodgson, Bree; Warren, Kimberly

doi:10.1016/j.rbmo.2017.10.039

Cited by 1 publication

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“…In future studies, it may also be feasible to streamline the technique even further to perform PGT for aneuploidy testing and PGT-M simultaneously in a single reaction. Options include the addition of primers with defined sequencing tags designed for mutation/HLA region after WGA using DOPlify 25 or, alternatively, taking an aliquot and performing PGT-M as above and adding this to the original WGA to get all results in one sequencing run. Mapping and counting of the sequencing reads are then performed, and the 24chromosome copy number plots are generated for ploidy assessment while bioinformatic identification of the gene/ HLA/SNP fragments enables the alleles to be scored.…”

Section: Scenariomentioning

confidence: 99%

Single-Molecule Sequencing

Rechitsky¹,

Kuliev²,

Ramon³

et al. 2020

The Journal of Molecular Diagnostics

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We investigated the potential of next-generation sequencing (NGS) as an alternative method for preimplantation genetic testing of monogenic disease (PGT-M) with human leukocyte antigen (HLA) matching and for noninvasive prenatal diagnosis follow-up. The case involved parents who were carriers of the Fanconi anemia complementation group G (FANCG) 260delG mutation. After clinical PGT using conventional short tandem repeat and mutation analysis, two euploid disease-free embryos were transferred, resulting in a twin pregnancy. Using the original embryo whole genome amplification products from 10 embryos, NGS confirmed the genotypes of the eight nontransferred embryos for both mutation status and HLA combination. NGS also confirmed that the two transferred embryos, which resulted in a twin pregnancy, were euploid, Fanconi disease free, and HLA matched to their sick sibling. At 15 weeks' gestation, noninvasive prenatal diagnosis of the maternal cell-free DNA determined fetal fractions of 14% and 6.6% for twins 1 and 2, respectively. The maternal plasma FANCG 260delG mutation ratio was measured at 46.2%, consistent with the presence of a carrier fetus and a normal fetus. These findings provide proof of concept that NGS has clinical utility as a safe and effective PGT-M method for embryo genotyping as well as more complex direct HLA matching. In addition, NGS can be used to confirm the original PGT-M and HLA matching embryo results in early pregnancy without the need for invasive prenatal diagnosis.

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Section: Scenariomentioning

confidence: 99%