PURPOSE: Preimplantation genetic testing for monogenic disorders (PGT-M) allows early diagnosis in embryos conceived in vitro. PGT-M helps to prevent known genetic disorders in affected families and ensures that pathogenic variants in the male or female partner are not passed on to offspring. The trend in genetic testing of embryos is to provide a comprehensive platform that enables robust and reliable testing for the causal pathogenic variant(s), as well as chromosomal abnormalities that commonly occur in embryos. In this study, we describe PGT protocol that allows direct mutation testing, haplotyping, and aneuploidy screening. METHODS: Described PGT protocol called OneGene PGT allows direct mutation testing, haplotyping, and aneuploidy screening using next-generation sequencing (NGS). Whole genome amplification product is combined with multiplex PCR used for SNP enrichment. Dedicated bioinformatic tool enables mapping, genotype calling, and haplotyping of informative SNP markers. A commercial software was used for aneuploidy calling. RESULTS: OneGenePGT has been implemented for seven of the most common monogenic disorders, representing approximately 30% of all PGT-M indications at our IVF centre. The technique has been thoroughly validated, focusing on direct pathogenic variant testing, haplotype identification, and chromosome abnormality detection. Validation results show full concordance with Sanger sequencing and karyomapping, which were used as reference methods. CONCLUSION: OneGene PGT is a comprehensive, robust, and cost-effective method that can be established for any gene of interest. The technique is particularly suitable for common monogenic diseases, which can be performed based on a universal laboratory protocol without the need for set-up or pre-testing.
- MeSH
- aneuploidie MeSH
- blastocysta patologie MeSH
- genetické testování metody MeSH
- lidé MeSH
- mutace genetika MeSH
- preimplantační diagnóza * metody MeSH
- těhotenství MeSH
- vysoce účinné nukleotidové sekvenování metody MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- těhotenství MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
PURPOSE: To evaluate the decline in transferable embryos in preimplantation genetic testing for aneuploidy (PGT-A) cycles due to (a) non-biopsable blastocyst quality, (b) failure of genetic analysis, (c) diagnosis of uniform numerical or structural chromosomal aberrations, and/or (d) chromosomal aberrations in mosaic constitution. METHODS: This retrospective multicenter study comprised outcomes of 1562 blastocysts originating from 363 controlled ovarian stimulation cycles, respectively, 226 IVF couples in the period between January 2016 and December 2018. Inclusion criteria were PGT-A cycles with trophectoderm biopsy (TB) and next generation sequencing (NGS). RESULTS: Out of 1562 blastocysts, 25.8% were lost due to non-biopsable and/or non-freezable embryo quality. In 10.3% of all biopsied blastocysts, genetic analysis failed. After exclusion of embryos with uniform or chromosomal aberrations in mosaic, only 18.1% of those originally yielded remained as diagnosed euploid embryos suitable for transfer. This translates into 50.4% of patients and 57.6% of stimulated cycles with no euploid embryo left for transfer. The risk that no transfer can take place rose significantly with a lower number of oocytes and with increasing maternal age. The chance for at least one euploid blastocyst/cycle in advanced maternal age (AMA)-patients was 33.3% compared to 52.1% in recurrent miscarriage (RM), 59.8% in recurrent implantation failure (RIF), and 60.0% in severe male factor (SMF). CONCLUSIONS: The present study demonstrates that PGT-A is accompanied by high embryo drop-out rates. IVF-practitioners should be aware that their patients run a high risk of ending up without any embryo suitable for transfer after (several) stimulation cycles, especially in AMA patients. Patients should be informed in detail about the frequency of inconclusive or mosaic results, with the associated risk of not having an euploid embryo available for transfer after PGT-A, as well as the high cost involved in this type of testing.
- MeSH
- aneuploidie MeSH
- blastocysta patologie MeSH
- genetické testování metody MeSH
- lidé MeSH
- preimplantační diagnóza * metody MeSH
- retrospektivní studie MeSH
- těhotenství MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- těhotenství MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- multicentrická studie MeSH
- MeSH
- fertilizace in vitro * metody MeSH
- kultivace embrya MeSH
- lidé MeSH
- preimplantační diagnóza * klasifikace MeSH
- těhotenství MeSH
- Check Tag
- lidé MeSH
- těhotenství MeSH
- ženské pohlaví MeSH
- Publikační typ
- přehledy MeSH
OBJECTIVE: Genomics Quality Assessment has provided external quality assessments (EQAs) for preimplantation genetic testing (PGT) for 12 years for eight monogenic diseases to identify sub-optimal PGT strategies, testing and reporting of results, which can be shared with the genomics community to aid optimised standards of PGT services for couples. METHOD: The EQAs were provided in two stages to mimic end-to-end protocols. Stage 1 involved DNA feasibility testing of a couple undergoing PGT and affected proband. Participants were required to report genotyping results and outline their embryo testing strategy. Lymphoblasts were distributed for mock embryo testing for stage 2. Submitted clinical reports and haplotyping results were assessed against peer-ratified criteria. Performance was monitored to identify poor performance. RESULTS: The most common testing methodology was short tandem repeat linkage analysis (59%); however, the adoption of single nucleotide polymorphism-based platforms was observed and a move from blastomere to trophectoderm testing. There was a variation in testing strategies, assigning marker informativity and understanding test limitations, some clinically unsafe. Critical errors were reported for genotyping and interpretation. CONCLUSION: EQA provides an overview of the standard of preimplantation genetic testing-M clinical testing and identifies areas of improvement for accurate detection of high-risk embryos.
- MeSH
- aneuploidie MeSH
- blastocysta MeSH
- genetické testování metody MeSH
- lidé MeSH
- preimplantační diagnóza * metody MeSH
- těhotenství MeSH
- Check Tag
- lidé MeSH
- těhotenství MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
Asistovaná reprodukce je nedílnou součástí léčby neplodnosti. Základní metodou je mimotělní oplození (IVF – fertilizace in vitro), běžně se používá i intracytoplazmatická injekce spermie do oocytu, kryokonzervace spermií, oocytů a embryí. Podíl embryí preimplantačně geneticky testovaných (PGT) na chromozomální aneuploidie nebo vady konkrétních genů stoupá. Významné je i použití darovaných oocytů, zejména z důvodu vyčerpání ovariální rezervy u žen starších 40 let. Efektivita léčby je vždy velmi zásadně závislá na věku ženy. Léčba asistovanou reprodukci je v Česku velmi dobře dostupná, stejně tak i záchrana plodnosti zmražením spermií/oocytů.
Assisted reproduction is an integral part of infertility treatment. The basic method is in vitro fertilisation (IVF), and intracytoplasmic sperm injection into the oocyte, cryopreservation of sperm, oocytes and embryos is also commonly used. The proportion of embryos tested with preimplantation genetic testing (PGT) for chromosomal aneuploidy or defects in specific genes is increasing. The use of donated oocytes is also significant, particularly because of the depletion of ovarian reserve in women over 40 years of age. The effectiveness of treatment is always very fundamentally dependent on the age of the woman. Assisted reproductive treatment is very well available in the Czech Republic, as is fertility preservation by sperm/oocyte freezing.
- MeSH
- asistovaná reprodukce * ekonomika zákonodárství a právo MeSH
- infertilita terapie MeSH
- lidé MeSH
- preimplantační diagnóza MeSH
- registrace MeSH
- věkové faktory MeSH
- Check Tag
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- přehledy MeSH
- Geografické názvy
- Česká republika MeSH
- MeSH
- asistovaná reprodukce klasifikace MeSH
- fertilizace in vitro klasifikace MeSH
- infertilita * terapie MeSH
- lidé MeSH
- preimplantační diagnóza klasifikace MeSH
- Check Tag
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- přehledy MeSH
Text reflektuje skutečnost, že preimplantační genetická diagnostika poskytuje naději na zdravé dítě i takovým párům, u kterých více pokusů o zdravého potomka selhalo. V rámci tohoto vyšetření embryí se však dozvídáme také informaci o pohlaví. Tento fakt by neměl být zneužíván k realizaci přání o narození chlapce, nebo děvčete. Nechtěná, přestože zcela zdravá embrya jsou totiž likvidována.
The manuscript reflects the fact that pre-implantation genetic diagnosis provides hope for a healthy child even for couples who have failed several times to have a healthy descendant. However, as part of this examination of the embryos, we also learn information about the sex. This fact should not be abused to realize the wish for the birth of a boy or a girl. Although completely healthy, unwanted embryos are disposed.
Text reflektuje skutečnost, že preimplantační genetická diagnostika poskytuje naději na zdravé dítě i takovým párům, u kterých více pokusů o zdravého potomka selhalo. V rámci tohoto vyšetření embryí se však dozvídáme také informaci o pohlaví. Tento fakt by neměl být zneužíván k realizaci přání o narození chlapce, nebo děvčete. Nechtěná, přestože zcela zdravá embrya jsou totiž likvidována.
The manuscript reflects the fact that pre-implantation genetic diagnosis provides hope for a healthy child even for couples who have failed several times to have a healthy descendant. However, as part of this examination of the embryos, we also learn information about the sex. This fact should not be abused to realize the wish for the birth of a boy or a girl. Although completely healthy, unwanted embryos are disposed.
- MeSH
- asistovaná reprodukce * dějiny ekonomika trendy MeSH
- infertilita diagnóza etiologie terapie MeSH
- lidé MeSH
- ovariální rezerva fyziologie MeSH
- počet spermií klasifikace MeSH
- preimplantační diagnóza metody MeSH
- reprodukční zdraví normy MeSH
- věkové faktory MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- rozhovory MeSH
