Simultaneous detection of genomic imbalance in patients receiving preimplantation genetic testing for monogenic diseases (PGT-M)

作者：Lin Yang, Yan Xu, Yan Xu, Jun Xia, Huijuan Yan, Chenhui Ding, Qianyu Shi, Yujing Wu, Ping Liu, Jiafu Pan, Yan‐Hong Zeng, Yanyan Zhang, Fang Chen, Hui Jiang, Yanwen Xu, Yanwen Xu, Wei Li, Canquan Zhou, Ya Gao · 发表于：Frontiers in Genetics · 年份：2022 · DOI：10.3389/fgene.2022.976131 · 被引用次数：4 · 研究领域：Prenatal Screening and Diagnostics、Genomic variations and chromosomal abnormalities、Chromosomal and Genetic Variations

Background: Preimplantation genetic test for monogenic disorders (PGT-M) has been used to select genetic disease-free embryos for implantation during in vitro fertilization (IVF) treatment. However, embryos tested by PGT-M have risks of harboring chromosomal aneuploidy. Hence, a universal method to detect monogenic diseases and genomic imbalances is required. Methods: Here, we report a novel PGT-A/M procedure allowing simultaneous detection of monogenic diseases and genomic imbalances in one experiment. Library was prepared in a special way that multiplex polymerase chain reaction (PCR) was integrated into the process of whole genome amplification. The resulting library was used for one-step low-pass whole genome sequencing (WGS) and high-depth target enrichment sequencing (TES). Results: The TAGs-seq PGT-A/M was first validated with genomic DNA (gDNA) and the multiple displacement amplification (MDA) products of a cell line. Over 90% of sequencing reads covered the whole-genome region with around 0.3–0.4 × depth, while around 5.4%–7.3% of reads covered target genes with >10000 × depth. Then, for clinical validation, 54 embryos from 8 women receiving PGT-M of β-thalassemia were tested by the TAGs-seq PGT-A/M. In each embryo, an average of 20.0 million reads with 0.3 × depth of the whole-genome region was analyzed for genomic imbalance, while an average of 0.9 million reads with 11260.0 × depth of the target gene HBB were analyzed for β-thalassemia. Eventually, 18 embry...