Robust Preimplantation Genetic Testing of Huntington Disease by Combined Triplet-Primed PCR Analysis of the HTT CAG Repeat and Multi-Microsatellite Haplotyping
| Autor: | Felicia S.H. Cheah, Mingjue Zhao, Gui Ping Phang, Arnold Sia Chye Tan, Anupriya Agarwal, Mulias Lian, Samuel S. Chong |
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| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
congenital hereditary and neonatal diseases and abnormalities Genetic testing Huntingtin lcsh:Medicine Locus (genetics) Fertilization in Vitro Biology Article 03 medical and health sciences 0302 clinical medicine mental disorders medicine Humans Allele lcsh:Science Genotyping Alleles Preimplantation Diagnosis Genetics Huntingtin Protein Multidisciplinary medicine.diagnostic_test lcsh:R Haplotype Pedigree Huntington Disease 030104 developmental biology Haplotypes Microsatellite lcsh:Q Single-Cell Analysis Trinucleotide Repeat Expansion Trinucleotide repeat expansion Multiplex Polymerase Chain Reaction 030217 neurology & neurosurgery Microsatellite Repeats Multilocus Sequence Typing |
| Zdroj: | Scientific Reports, Vol 9, Iss 1, Pp 1-11 (2019) Scientific Reports |
| ISSN: | 2045-2322 |
| Popis: | Huntington disease (HD) is a lethal neurodegenerative disorder caused by expansion of a CAG repeat within the huntingtin (HTT) gene. Disease prevention can be facilitated by preimplantation genetic testing for this monogenic disorder (PGT-M). We developed a strategy for HD PGT-M, involving whole genome amplification (WGA) followed by combined triplet-primed PCR (TP-PCR) for HTT CAG repeat expansion detection and multi-microsatellite marker genotyping for disease haplotype phasing. The strategy was validated and tested pre-clinically in a simulated PGT-M case before clinical application in five cycles of a PGT-M case. The assay reliably and correctly diagnosed all embryos, even where allele dropout (ADO) occurred at the HTT CAG repeat locus or at one or more linked markers. Ten of the 27 embryos analyzed were diagnosed as unaffected. Four embryo transfers were performed, two of which involved fresh cycle double embryo transfers and two were frozen-thawed single embryo transfers. Pregnancies were achieved from each of the frozen-thawed single embryo transfers and confirmed to be unaffected by amniocentesis, culminating in live births at term. This strategy enhances diagnostic confidence for PGT-M of HD and can also be employed in situations where disease haplotype phase cannot be established prior to the start of PGT-M. |
| Databáze: | OpenAIRE |
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