Evaluation of targeted next-generation sequencing-based preimplantation genetic diagnosis of monogenic disease

Objective: To investigate the applicability of next-generation sequencing (NGS) to preimplantation genetic diagnosis (PGD); to evaluate semiconductor-based NGS for genetic analysis of human embryos. Design: Blinded. Setting: Academic center for reproductive medicine. Patient(s): Six couples at risk of transmitting single-gene disorders to their offspring. Intervention(s): None. Main Outcome Measure(s): Embryonic genotype consistency of NGS with two independent conventional methods of PGD. Result(s): NGS provided 100% equivalent PGD diagnoses of compound point mutations and small deletions and insertions compared with both reference laboratory-and internally developed quantitative polymerase chain reaction (qPCR)-based analyses. Furthermore, NGS single-gene disorder screening could be performed in parallel with qPCR-based comprehensive chromosome screening. Conclusion(s): NGS can provide blastocyst PGD results with a high level of consistency with established methodologies. This study and its design could serve as a model for further development of this important and emerging technology. (Fertil Steril (R) 2013; 99:1377-84. (C) 2013 by American Society for Reproductive Medicine.)