Validating a rapid, real-time, PCR-based direct mutation detection assay for preimplantation genetic diagnosis
Journal
Gene
Journal Volume
299
Journal Issue
305
Pages
299-305
Date Issued
2014
Author(s)
HSIN-FU CHEN
Chang S.-P.
Wu S.-H.
Lin W.-H.
Lee Y.-C.
Ma G.-C.
Ginsberg N.A.
You E.-M.
Tsai F.-P.
MING CHEN
Abstract
Although co-amplification of polymorphic microsatellite markers is the current gold standard for preimplantation genetic diagnosis (PGD) of single-gene disorders (SGD), this approach can be hampered by the lack of availability of informative markers. We recently (2011) devised a novel in-house assay for PGD of aromatic l-amino acid decarboxylase deficiency, based on an amplification refractory mutation system and quantitative PCR (ARMS-qPCR). The objective of the present study was to verify ARMS-qPCR in a cohort of 20 PGD cycles with a diverse group of SGDs (15 couples at risk for 10 SGDs). Day-3 cleavage-stage embryos were subjected to biopsy and genotyping, followed by fresh embryo transfer (FET). The diagnostic rate was 82.9%; unaffected live births were achieved in 9 of 20 FET cycles (45%), with only one false negative (among 54 transferred embryos). Overall, the ARMS-qPCR had frequent allele-dropout (ADO), rendering it inappropriate as the sole diagnostic method (despite a favorable live-birth rate). Regardless, it has the potential to complement the current gold-standard methodology, especially when trophectoderm biopsy becomes a preferred option and genotyping needs to be timely enough to enable FET. ? 2014 Elsevier B.V.
SDGs
Other Subjects
acute intermittent porphyria; adult; amino acid deficiency; article; beta thalassemia; biopsy technique; centronuclear myopathy; clinical article; cohort analysis; controlled study; diagnostic accuracy; Duchenne muscular dystrophy; embryo; embryo transfer; female; gene amplification; gene frequency; gene mutation; genetic analysis; genotype; gold standard; hearing impairment; hereditary motor sensory neuropathy; human; kidney polycystic disease; live birth; preimplantation genetic diagnosis; prenatal diagnosis; priority journal; quantitative analysis; real time polymerase chain reaction; retrospective study; risk factor; spinal muscular atrophy; ARMS-qPCR; Cleavage-stage embryo biopsy; Fresh embryo transfer (FET); Pre-implantation genetic diagnosis (PGD); Trophectoderm biopsy; Adult; Embryo Transfer; Embryo, Mammalian; Female; Genetic Predisposition to Disease; Genetic Testing; Genotyping Techniques; Humans; Male; Mutation; Pregnancy; Preimplantation Diagnosis; Real-Time Polymerase Chain Reaction
Type
journal article