Development of Preimplantation Genetic Testing for Monogenic Reference Materials Based on Next Generation Sequencing

Preimplantation genetic testing for monogenic disorders (PGT-M) is available for detecting a majority of serious genetic conditions over the past 20 years. However, there are still lack of reference materials to validate the test performance during test development and quality control in PGT-M cycles. In this study, PGT-M reference materials containing 12 gDNA reference materials and 4 simulated embryo reference materials for thalassemia testing were successfully established and well characterized. Next generation sequencing was performed on 12 gDNA samples and 4 simulated embryo samples and the genotypes and haplotypes of all 16 PGT-M reference materials for thalassemia showed no discordance from 4 labs with varies testing workflows. And the well-characterized PGT-M reference materials achieved good stability even after 3 years of storage. Therefore, the PGT-M reference materials for thalassemia established in this work will enable the standardization and high accuracy of PGT-M in clinical cases. This pioneer methodology can be easily expanded to develop PGT-M reference materials for other monogenic disease.