OneGene PGT: comprehensive preimplantation genetic testing method utilizing next-generation sequencing

• Published in: Journal of assisted reproduction and genetics 2024-01, Vol.41 (1), p.185-192
• Main Authors: Hornak, Miroslav, Bezdekova, Katerina, Kubicek, David, Navratil, Rostislav, Hola, Veronika, Balcova, Maria, Bohmova, Magdalena, Weisova, Katerina, Vesela, Katerina
• Format: Article
• Language: English
• Subjects: Aneuploidy; Blastocyst - pathology; Chromosome aberrations; Embryos; Female; Gene mapping; Genetic disorders; Genetic screening; Genetic testing; Genetic Testing - methods; Genetics; Genomes; Genotypes; Gynecology; Haplotypes; High-Throughput Nucleotide Sequencing - methods; Human Genetics; Humans; Male; Medicine; Medicine & Public Health; Mutation; Mutation - genetics; Next-generation sequencing; Pregnancy; Preimplantation Diagnosis - methods; Reproductive Medicine; Single-nucleotide polymorphism
• ISSN: 1058-0468; 1573-7330; 1573-7330
• DOI: 10.1007/s10815-023-02998-3

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.