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Precise Exome Analysis Of Blastocyst Biopsy Scale Samples Using Primary Template-Directed Amplification

Samitova, A.; Belova, V.; Vasiliadis, I.; Repinskaia, Z.; Gorodnicheva, T.; Romanov, E.; Pogosyan, M.; Gaysin, E.; Nazarenko, T.; Rebrikov, D.; Korostin, D.

2024-11-03 genetics
10.1101/2024.10.29.620888 bioRxiv
Show abstract

This study evaluates primary template-directed amplification (PTA) for whole exome sequencing (WES) on small fibroblast cell groups, mimicking the limited cell quantities typical of trophectoderm embryo biopsies. PTAs consistent amplification reduces allelic dropout (ADO) and impoves uniform coverage, overcoming challenges associated with conventional methods such as multiple displacement amplification (MDA). Using fibroblast samples alongside well-characterised genomic references (E701, NA12878), we benchmarked PTA-WES, achieving 97.5% target region coverage at 10x, meeting American College of Medical Genetics and Genomics (ACMG) standards. Preliminary results from embryo biopsies sequenced with PTA-WES showed a median coverage of 102x, significantly improving upon the variability and coverage gaps observed in MDA-WES. The findings support PTAs potential to enhance the clinical applicability of WES for preimplantation genetic testing for monogenic disorders (PGT-M), expanding capabilities to detect inherited and de novo mutations in embryos. Further optimisation and variant detection analyses are planned to evaluate PTAs robustness for routine clinical use.

Published in BMC Genomics (predicted rank #2) · training set

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